Triazaspiro compounds useful for treating or preventing pain

ABSTRACT

Triazaspiro Compounds, compositions comprising a Triazaspiro Compound, methods for treating or preventing pain in an animal comprising administering to an animal in need thereof an effective amount of a Triazaspiro Compound and methods for stimulating opioid-receptor function in a cell comprising contacting a cell capable of expressing an opioid receptor with an effective amount of a Triazaspiro Compound are disclosed.

This application claims the benefit of U.S. provisional application No.60/384,807, filed May 31, 2002, and of U.S. provisional application No.60/460,219, filed Apr. 3, 2003, the disclosure of each of which isincoporated by reference in its entirety.

1. FIELD OF THE INVENTION

The present invention relates to Triazaspiro Compounds, compositionscomprising a Triazaspiro Compound and methods for preventing or treatingpain in an animal in need thereof comprising administering to the animalan effective amount of a Triazaspiro Compound.

2. BACKGROUND OF THE INVENTION

Pain is the most common symptom for which patients seek medical adviceand treatment. Pain can be acute or chronic. While acute pain is usuallyself-limited, chronic pain can persist for 3 months or longer and leadto significant changes in a patient's personality, lifestyle, functionalability or overall quality of life (K. M. Foley, Pain, in Cecil Textbookof Medicine 100-107 (J. C. Bennett and F. Plum eds., 20th ed. 1996).

Pain has been traditionally managed by administering a non-opioidanalgesic, such as acetylsalicylic acid, choline magnesiumtrisalicylate, acetaminophen, ibuprofen, fenoprofen, diflusinal andnaproxen; or an opioid analgesic, including morphine, hydromorphone,methadone, levorphanol, fentanyl, oxycodone and oxymorphone. Id.

U.S. Pat. No. 3,238,216 to Janssen et al. discloses particularspirocompounds allegedly useful as neuroleptic, including analgesic,agents.

International Publication No. WO 99/45011 by Janssen Pharmaceutica N.V.discloses particular spirocompounds allegedly useful for the treatmentof pain.

U.S. Pat. No. 6,277,991 to Hohlweg et al. discloses particularspirocompounds allegedly useful for the treatment of migraine headache.

Traditional non-opioid analgesics exert their pharmacological activityonce they have passed through the blood-brain barrier. But thisblood-brain barrier passage can lead to many undesirable central nervoussystem-mediated side effects, such as respiratory depression, increaseddrug-abuse potential, increased drug tolerance, increased drugdependence, constipation and unwanted euphoria.

U.S. Pat. No. 6,362,203 to Mogi et al. discloses particular4-hydroxy-4-phenylpiperidine compounds allegedly useful as peripheralanalgesic agents.

There remains a clear need in the art for new drugs useful for treatingor preventing pain and that reduce or avoid one or more side effectsassociated with traditional therapy for treating pain.

Citation of any reference in Section 2 of this application is not to beconstrued as an admission that such reference is prior art to thepresent application.

3. SUMMARY OF THE INVENTION

The present invention encompasses compounds having the formula (Ia):

and pharmaceutically acceptable salts thereof, wherein:

R¹ is hydrogen, —COOH, —COOR³, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl),—C(O)N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —CN, —N(H)S(O)₂(C₁-C₄ alkyl), or

R² is —COOH, —COOR³ or

R³ is —(C₁-C₆ alkyl), benzyl, phenyl, or —(C₃-C₆ cycloalkyl);

n is 0 when R¹ is hydrogen, and n is an integer ranging from 1 to 4 whenR¹ is other than hydrogen; and

m is an integer ranging from 0 to 4.

The present invention also encompasses compounds having the formula(Ib):

and pharmaceutically acceptable salts thereof, wherein:

R¹ is —COOH, —COOR³, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), —C(O)N(C₁-C₄alkyl)(C₁-C₄ alkyl), —CN, —N(H)S(O)₂(C₁-C₄ alkyl), or

R² is —COOH, —COOR³, —C(O)N(CH₃)₂ or

R³ is —(C₁-C₆ alkyl), benzyl, phenyl, or —(C₃-C₆ cycloalkyl);

n is an integer ranging from 1 to 4; and

m is an integer ranging from 0 to 4.

The present invention also encompasses compounds having the formula(Ic):

and pharmaceutically acceptable salts thereof, wherein:

R¹ is —COOH, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), —C(O)N(C₁-C₄ alkyl)(C₁-C₄alkyl), —N(H)S(O)₂(C₁-C₄ alkyl), or

n is an integer ranging from 1 to 4.

A Triazaspiro Compound is useful for treating or preventing pain in ananimal.

The invention also relates to compositions comprising an effetive amountof a Triazaspiro Compound and a pharmaceutically acceptable carrier orexcipient. The present compositions are useful for treating orpreventing pain in an animal.

The invention also relates to kits comprising a container containing aTriazaspiro Compound, and instructions for its use.

The invention further relates to methods for preventing pain in ananimal, comprising administering to an animal in need thereof aneffective amount of a Triazaspiro Compound.

The invention further relates to methods for treating pain in an animal,comprising administering to an animal in need thereof an effectiveamount of a Triazaspiro Compound.

The invention still further relates to methods for stimulatingopioid-receptor function in a cell, comprising contacting a cell capableof expressing an opioid receptor with an effective amount of aTriazaspiro Compound.

In a still further embodiment, the present invention is directed towarda method for preparing a composition, comprising the step of admixing aTriazaspiro Compound and a pharmaceutically acceptable carrier orexcipient.

The present invention may be understood more fully by reference to thefollowing detailed description and illustrative examples, which areintended to exemplify non-limiting embodiments of the invention.

4. DETAILED DESCRIPTION OF THE INVENTION 4.1 Definitions

As used herein, the terms used above having following meaning:

A “Triazaspiro Compound” is a compound of formula (Ia), formula (Ib) orformula (Ic), a compound set forth in Section 4.3 of this application,or a pharmaceutically acceptable salt of any of the above.

“—C₁-C₄ alkyl” means a straight or branched non-cyclic hydrocarbon chainhaving from 1 to 4 carbon atoms. Representative straight chain —C₁-C₄alkyls include -methyl, -ethyl, -n-propyl, and -n-butyl. Representativebranched chain —C₁-C₄ alkyls include -isopropyl, -sec-butyl, -isobutyl,and -tert-butyl.

“—C₁-C₆ alkyl” means a straight or branched non-cyclic hydrocarbon chainhaving from 1 to 6 carbon atoms. Representative straight chain —C₁-C₆alkyls include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyland-n-hexyl. Representative branched chain —C₁-C₆ alkyls include-isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, -neopentyl,1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl,1,2-dimethylpropyl, 1-methylpentyl, 2-methylpentyl, 3-methylpentyl,4-methylpentyl, 1-ethylbutyl, 2-ethylbutyl, 3-ethylbutyl,1,1-dimethtylbutyl, 1,2-dimethylbutyl, 1,3-dimethylbutyl,2,2-dimethylbutyl, 2,3-dimethylbutyl and 3,3-dimethylbutyl.

“—C₃-C₆ cycloalkyl” means a saturated cyclic hydrocarbon having from 3to 6 carbon atoms. Representative —C₃-C₆ cycloalkyls are -cyclopropyl,-cyclobutyl, -cyclopentyl and cyclohexyl.

The term “animal,” includes, but is not limited to, a cow, monkey,horse, sheep, pig, chicken, turkey, quail, cat, dog, mouse, rat, rabbit,guinea pig and human.

The phrase “pharmaceutically acceptable salt,” as used herein, is a saltformed from an acid and the basic nitrogen group of a TriazaspiroCompound. Such salts include, but are not limited, to sulfate, citrate,acetate, oxalate, chloride, bromide, iodide, nitrate, bisulfate,phosphate, acid phosphate, isonicotinate, lactate, salicylate, acidcitrate, tartrate, oleate, tannate, pantothenate, bitartrate, ascorbate,succinate, maleate, gentisinate, fumarate, gluconate, glucaronate,saccharate, formate, benzoate, glutamate, methanesulfonate,ethanesulfonate, benzenesulfonate, p-toluenesulfonate, and pamoate(i.e., 1,1′-methylene-bis-(2-hydroxy-3-naphthoate)) salts. The term“pharmaceutically acceptable salt” also refers to a salt of aTriazaspiro Compound having an acidic functional group, such as acarboxylic acid functional group, and a pharmaceutically acceptableinorganic or organic base. Suitable bases include, but are not limitedto, hydroxides of alkali metals such as sodium, potassium and lithium;hydroxides of alkaline earth metal such as calcium and magnesium;hydroxides of other metals, such as aluminum and zinc; ammonia; andorganic amines, such as unsubstituted or hydroxy-substituted mono-, di-,or trialkylamines; dicyclohexylamine; tributylamine; pyridine;N-methyl-N-ethylamine; diethylamine; triethylamine; mono-, bis-, ortris-(2-hydroxy-lower alkyl amines), such as mono-, bis-, ortris-(2-hydroxyethyl)amine, 2-hydroxy-tert-butylamine, ortris-(hydroxymethyl)methylamine, N,N,-di-lower alkyl-N-(hydroxy loweralkyl)-amines, such as N,N,-dimethyl-N-(2-hydroxyethyl)amine, ortri-(2-hydroxyethyl)amine; N-methyl-D-glucamine; and amino acids such asarginine, lysine, and the like.

The phrase “treatment of” and “treating” pain includes the ameliorationor cessation of pain.

The phrase “prevention of” and “preventing” pain includes the avoidanceof the onset of pain.

The phrase “opioid receptor” means a δ-opioid receptor, a κ-opioidreceptor, a μ-opioid receptor or an ORL-1 receptor.

The phrase “effective amount” when used in connection with a TriazaspiroCompound means an amount effective for (a) treating or preventing paint,or (b) stimulating opioid receptor function in a cell.

The phrase “effective amount” when used in connection with anothertherapeutic agents means an amount for providing the therapeutic effectof the therapeutic agent.

When a first group is “substituted with one or more” second groups, oneor more hydrogen atoms of the first group is replaced with acorresponding number of second groups. When the number of second groupsis two or greater, the second groups can be the same or different. Inone embodiment, the number of second groups is 1 or 2. In anotherembodiment, the number of second groups is 1.

4.2 The Triazaspiro Compounds of Formulas (Ia)-(Ic)

4.2.1 The Triazaspiro Compounds of Formula (Ia)

As stated above, the present invention encompasses Triazaspiro Compoundshaving the formula (Ia):

and pharmaceutically acceptable salts thereof, wherein:

R¹ is hydrogen, —COOH, —COOR³, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl),—C(O)N(C₁-C₄ alkyl)(C₁-C₄ alkyl), —CN, —N(H)S(O)₂(C₁-C₄ alkyl), or

R² is —COOH, —COOR³ or

R³ is —(C₁-C₆ alkyl), benzyl, phenyl, or —(C₃-C₆ cycloalkyl);

n is 0 when R¹ is hydrogen, and n is an integer ranging from 1 to 4 whenR¹ is other than hydrogen; and

m is an integer ranging from 0 to 4.

In one embodiment, the Triazaspiro Compounds of formula (Ia) are thosewherein R¹ is hydrogen and n is 0.

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose whrein R¹ is other than hydrogen and n is an integer ranging from1 to 4.

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose wherein R¹ is —COOH or —COOR³, and R³ is —(C₁-C₆ alkyl), benzyl,phenyl, or —(C₃-C₆ cycloalkyl). In another embodiment, R³ is —CH₃.

In another embodiment, the Triazaspiro Compounds of of formula (Ia) arethose wherein R¹ is —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl) or —C(O)N(C₁-C₄alkyl)(C₁-C₄ alkyl). In another embodiment, R¹ is —C(O)NH(CH₃) or—C(O)N(CH₃)₂.

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose wherein R¹ is —CN.

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose wherein R¹ is —N(H)S(O)₂(C₁-C₄ alkyl). In another embodiment, R¹is —N(H)S(O)₂(CH₃).

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose wherein R¹ is

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose wherein R² is —COOH or —COOR³, and R³ is —(C₁-C₆ alkyl), benzyl,phenyl, or —(C₃-C₆ cycloalkyl). In another embodiment, R³ is —CH₃.

In another embodiment, t the Triazaspiro Compounds of formula (Ia) arethose wherein R² is

In another embodiment, m is 0 or 1. In another embodiment, m is 0.

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose wherein R¹ is H, n is 0, and R² is —COOR³, and R³ is —(C₁-C₆alkyl), benzyl, phenyl, or —(C₃-C₆ cycloalkyl). In another embodiment,R³ is —CH₃.

In another embodiment, the Triazaspiro Compounds of formula (Ia) arethose wherein R¹ is H, n is 0, R² is

and m is 0.

Illustrative Triazaspiro Compounds of formula (Ia) are:

-   4-(4-Oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-8-yl)-2,2-diphenyl-butyric    acid methyl ester;-   4-(4-Oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-8-yl)-2,2-diphenyl-butyric    acid;-   8-[3,3-Diphenyl-3-(1H-tetrazol-5-yl)-propyl]-1-phenyl-1,3,8-triaza-spiro[4.5]decan-4-one;

and pharmaceutically acceptable salts thereof.

4.2.2 The Triazaspiro Compounds of Formula (Ib)

The present invention also encompasses Triazaspiro Compounds having theformula (Ib):

and pharmaceutically acceptable salts thereof, wherein:

R¹ is —COOH, —COOR³, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), —C(O)N(C₁-C₄alkyl)(C₁-C₄ alkyl), —CN, —N(H)S(O)₂(C₁-C₄ alkyl), or

R² is —COOH, —COOR³, —C(O)N(CH₃)₂, or

R³ is —(C₁-C₆ alkyl), benzyl, phenyl, or —(C₃-C₆ cycloalkyl);

n is an integer ranging from 1 to 4; and

m is an integer ranging from 0 to 4.

In one embodiment, the Triazaspiro Compounds of formula (Ib) are thosewherein R¹ is —COOH or —COOR³, and R³ is —(C₁-C₆ alkyl), benzyl, phenyl,or —(C₃-C₆ cycloalkyl). In one embodiment, R³ is —CH₃.

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R¹ is —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), or —C(O)N(C₁-C₄alkyl)(C₁-C₄ alkyl). In another embodiment, R¹ is —C(O)NH(CH₃) orC(O)N(CH₃)₂.

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R¹ is —CN.

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R¹ is —N(H)S(O)₂(C₁-C₄ alkyl). In antoher embodiment, R¹is N(H)S(O)₂(CH₃).

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R¹ is

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R² is —COOH or —COOR³, and R³ is —(C₁-C₆ alkyl), benzyl,phenyl, or —(C₃-C₆ cycloalkyl). In another embodiment, R³ is —CH₃.

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R² is —C(O)N(CH₃)₂.

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R² is

and m is an integer ranging from 0 to 4. In another embodiment, m is 0or 1. In another embodiment, m is 0.

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R¹ is —COOH and R² is —C(O)N(CH₃)₂.

In another embodiment, the Triazaspiro Compounds of formula (Ib) arethose wherein R¹ is —COOR³, R² is —C(O)N(CH₃)₂ and R³ is —(C₁-C₆ alkyl),benzyl, phenyl, or —(C₃-C₆ cycloalkyl). In another embodiment, R³ is—CH₃.

Illustrative Triazaspiro Compounds of formula (Ib) are:

-   5-[8-(3-Dimethylcarbamoyl-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-pentanoic    acid;-   [8-(3-Dimethylcarbamoyl-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-acetic    acid methyl ester;-   3-[8-(3-Dimethylcarbamoyl-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-propionic    acid ethyl ester;-   3-[8-(3-Dimethylcarbamoyl-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-propionic    acid; and    pharmaceutically acceptable salts thereof.    4.2.3 The Triazaspiro Compounds of Formula (Ic)

The present invention also encompasses Triazaspiro Compounds having theformula (Ic):

and pharmaceutically acceptable salts thereof, wherein:

R¹ is —COOH, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), —C(O)N(C₁-C₄ alkyl)(C₁-C₄alkyl), —N(H)S(O)₂(C₁-C₄ alkyl), or

n is an integer ranging from 1 to 4.

In one embodiment, the Triazaspiro Compounds of formula (Ic) are thosewherein R¹ is —COOH.

In another embodiment, the Triazaspiro Compounds of formula (Ic) arethose wherein R¹ is —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl) or —C(O)N(C₁-C₄alkyl)(C₁-C₄ alkyl). In another embodiment, R¹ is —C(O)NH(CH₃) or—C(O)N(CH₃)₂.

In another embodiment, the Triazaspiro Compounds of formula (Ic) arethose wherein R¹ is —N(H)S(O)₂(C₁-C₄ alkyl). In another embodiment, R¹is —NHS(O)₂(CH₃).

In another embodiment, the Triazaspiro Compounds of formula (Ic) arethose wherein R¹ is

In another embodiment, n is 0, 1, or 2. h another embodiment, n is 0or 1. In another embodiment, n is 2.

Illustrative compounds of formula (Ic) are:

-   [8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-acetic    acid;-   N-{2-[8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-ethyl}-methanesulfonamide;-   2-[8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-acetamide;-   3-[8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-propionic    acid;-   5-[8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-pentanoic    acid;-   8-(3,3-diphenyl-propyl)-1-phenyl-3-(1H-tetrazol-5-ylmethyl)-1,3,8-triaza-spiro[4.5]decan-4-one;-   8-(3,3-Diphenyl-propyl)-1-phenyl-3-[2-(1H-tetrazol-5-yl)-ethyl]-1,3,8-triaza-spiro[4.5]decan-4-one;    and pharmaceutically acceptable salts thereof.

4.3 Other Triazaspiro Compounds

Other Triazaspiro Compounds are:

-   4-(4-Oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-8-yl)-2,2-diphenyl-butyronitrile;-   N,N-Dimethyl-4-(4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-8-yl)-2,2-diphenyl-butyramide;-   [8-(3-Cyano-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-acetic    acid;-   [8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-acetic    acid ethyl ester;-   [8-(3-Cyano-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-acetic    acid ethyl ester;-   3-[8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-propionitrile;-   4-[8-(3,3-Diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-butyronitrile;-   5-[8-(3-Cyano-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-pentanoic    acid ethyl ester;-   5-[8-(3-Cyano-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-pentanoic    acid;-   3-[8-(3-Cyano-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-propionic    acid;-   3-[8-(3-cyano-3,3-diphenyl-propyl)-4-oxo-1-phenyl-1,3,8-triaza-spiro[4.5]dec-3-yl]-propionic    acid ethyl ester;    and pharmaceutically acceptable salts thereof.

4.4 Methods for Making the Triazaspiro Compounds

The Triazaspiro Compounds can be made using conventional organicsyntheses and/or by the following illustrative methods:

The Triazaspiro Compounds wherein R¹ is H, n is 0, and R² is —COOR³(wherein R³ is defined above) can be prepared by reacting Compound Awith Compound B wherein R² is —COOR³ (wherein R³ is defined above) and Xis a leaving group such as a benzenesulfonate, 4-methylbenzenesulfonate,4-bromobenzenesulfonate, 4-nitrobenzenesulfonate, methanesulfonate,trifluoromethanesulfonate, or halogen in the presence of an organicbase, such as pyridine, 4-dimethylpyridine, triethylamine, ordiisopropylethylamine (DIEA), in an aprotic solvent to provide CompoundC wherein R² is —COOR³ (wherein R³ is defined above). In one embodimentX is I or Br. Suitable aprotic solvents include, but are not limited to,acetonitrile, dimethylsulfoxide (DMSO), dimethylformamide (DMF),dichloromethane (DCM), 1,2-dichloroethane, tetrahydrofuran (THF),diethyl ether, ligroin, pentane, hexane, and dioxane. In one embodimentthe aprotic solvent is acetonitrile.

The Triazaspiro Compounds wherein R¹ is H, n is 0, and R² is —COOH canbe prepared by hydrolyzing Compound C wherein R² is —COOR³ (wherein R³is defined above). The hydrolysis can be performed using an excess of anaqueous base, such as about 0.01 to about 1 N alkali metal hydroxide andthen acidifying the hydrolysis product. The acidifying can be performedusing about 0.01 to 3 N acid. In one embodiment the acid HCl. In oneembodiment the alkali metal hydroxide is potassium hydroxide.

The Triazaspiro Compounds wherein R¹ is H, n is 0, and R² is

(wherein m is defined above) can be prepared by reacting Compound A andCompound B wherein R² is —(CH₂)_(m)CN to obtain Compound C wherein R² is—(CH₂)_(m)CN. Compound C wherein R² is —(CH₂)_(m)CN is then reacted withtrimethylsilylazide (“TMSN_(3”)) in the presence of tin oxide accordingto the procedure described in S. J. Wittenberg et al., J. Org. Chem.58;4139-4141 (1993).

The Triazaspiro Compound wherein R¹ is H, n is 0, and R² is —C(O)N(CH₃)₂(Compound AB) can be prepared as described below:

Compound B(1) (obtainable by hydrolyzing Compound B wherein R² is—COOR³) is converted to an acid chloride using thionyl chloride oroxaloyl chloride to provide Compound D. Methods for obtaining acidchlorides are well known to those skilled in the art. Compound D is thenreacted with about 1 to about 10 molar eq. of dimethylamine and analkali metal carbonate, an alkali metal bicarbonate, or an alkalineearth carbonate to provide Compound E. One eq. of Compound E is thenreacted with about 1 eq. of Compound A in the presence of an alkalimetal carbonate, an alkali metal bicarbonate, or an alkaline earthcarbonate in an aprotic solvent defined above to provide Compound AB. Inone embodiment the alkali metal carbonate is Na₂CO₃. In anotherembodiment the aprotic solvent is DMF. Typically the reaction ofCompound A and Compound E is conducted at a temperature of from aboutroom temperature to about 100° C.

Compound C wherein R² is H can be prepared by reacting Compound A withCompound B wherein R² is H, and X is a leaving group such as abenzenesulfonate, 4-methylbenzenesulfonate, 4-bromobenzenesulfonate,4-nitrobenzenesulfonate, methanesulfonate, trifluoromethanesulfonate, orhalogen in the presence of an organic base, such as pyridine,4-dimethylpyridine, triethylamine, or diisopropylethylamine (DIEA), inan aprotic solvent. In one embodiment X is I or Br. Suitable aproticsolvents include, but are not limited to, acetonitrile,dimethylsulfoxide (DMSO), dimethylformamide (DMF), dichloromethane(DCM), 1,2-dichloroethane, tetrahydrofuran (THF), diethyl ether,ligroin, pentane, hexane, and dioxane. In one embodiment the aproticsolvent is acetonitrile.

The Triazaspiro Compounds wherein R¹ is other than H and n is an integerranging from 1 to 4 can be prepared by reacting Compound C wherein R² isH, —COOR³, —C(O)N(CH₃)₂, or

(wherein R₃ and m are defined above), with a compound of general formulaR¹(CH₂)_(n)—X wherein R¹ is other than H and is defined above, n is aninteger ranging from 1 to 4 and X is a leaving group such as abenzenesulfonate, 4-methylbenzenesulfonate, 4-bromobenzenesulfonate,4-nitrobenzenesulfonate, methanesulfonate, trifluoromethanesulfonate, orhalogen in the presence of a strong base such as lithiumdiisopropylamide or sodium hydride and an aprotic solvent such as THF,dioxane, or DMF. In one embodiment X is a halogen. In another embodimentX is I or Br. In another embodiment the base is NaH. In anotherembodiment the solvent is DMF.

The Triazaspiro Compounds wherein R¹ is other than H and R² is —COOH canbe prepared by hydrolyzing the Triazaspiro Compounds wherein R¹ is otherthan H and R² is —COOR³ (wherein R³ is defined above). The hydrolysiscan be performed using an excess of an aqueous base, such as about 0.01to about 1 N alkali metal hydroxide and then acidifying the hydrolysisproduct. The acidifying can be performed using about 0.01 to 3 N acid.In one embodiment the acid HCl. In one embodiment the alkali metalhydroxide is potassium hydroxide.

Compound A is commercially available from Sigma-Aldrich, St. Louis, Mo.(www.sigma-aldrich.com).

Compound B wherein R² is —COOH and X is Br is commercially availablefrom Sigma-Aldrich, St. Louis, Mo. (www.sigma-aldrich.com).

Compound B wherein R² is —COOR³ is obtainable by esterifying Compound Bwherein R² is —COOH (or a derivative thereof) and X is Br. Methods foresterifying carboxylic acids and derivatives thereof are well known tothose skilled in the art (See, e.g., J. March, Advanced OrganicChemistry, Reaction Mechanisms and Structure 392-396 and 400 (4^(th) ed.1992)).

Compound B wherein R² is —CN and X is Br is commercially available fromSigma-Aldrich, St. Louis, Mo. (www.sigma-aldrich.com).

Compound B wherein R² is —(CH₂)_(m)CN, m is an integer ranging from 1 to4 and X is Br is commercially available Sigma-Aldrich, St. Louis, Mo.(www.sigma-aldrich.com).

Compounds of general formula R¹ (CH₂)_(n)—X wherein R¹ is —COOH, X is ahalogen, and n is an integer ranging from 1 to 4 are commerciallyavailable from Sigma-Aldrich, St. Louis, Mo. (www.sigma-aldrich.com).

Compounds of general formula R¹(CH₂)_(n)—X wherein R¹ is —COOR³ (whereinR₃ is defined above), X is a halogen, and n is an integer ranging from 1to 4 are obtainable by esterifying compounds of general formulaR¹(CH₂)_(n)—X wherein R¹ is —COOH (or a derivative thereof). Methods foresterifying carboxylic acids or derivatives thereof are well known tothose skilled in the art (See, e.g., J. March, Advanced OrganicChemistry, Reaction Mechanisms and Structure 392-396 and 400 (4^(th) ed.1992)).

Compounds of general formula R¹(CH₂)_(n)—X wherein R¹ is —C(O)NH₂,—C(O)NH(C₁-C₄ alkyl) or —C(O)N(C₁-C₄ alkyl)(C₁-C₄ alkyl); X is ahalogen; and n is an integer ranging from 1 to 4 are obtainable byaminating a compound of general formula R¹(CH₂)_(n)—X wherein R¹ is—COOH (or a derivative thereof). Methods for aminating carboxylic acidsor derivatives thereof are well known to those skilled in the art (See,e.g., J. March, Advanced Organic Chemistry, Reaction Mechanisms andStructure 417-424 (4^(th) ed. 1992)).

Compounds of general formula R¹(CH₂)_(n)—X wherein R¹ is—N(H)S(O)₂(C₁-C₄ alkyl), X is a halogen, and n is an integer rangingfrom 1 to 4 can be obtained by reacting an amine of formulaNH₂—(CH₂)_(n)—X, wherein X is a halogen, and n is an integer rangingfrom 1 to 4 (commercially available from Sigma-Aldrich, St. Louis, Mo.(www.sigma-aldrich.com) with a sulfonyl chloride of formulaCl—S(O)₂(C₁-C₄ alkyl). The sulfonyl halides are commercially availableor can be made by methods well known to those skilled in the art (See,e.g., J. March, Advanced Organic Chemistry, Reaction Mechanisms andStructure, 499 (4^(th) ed. 1992)).

Compounds of general formula R¹(CH₂)_(n)—X wherein R¹ is —CN, X is ahalogen, and n is an integer ranging from 1 to 4 are commerciallyavailable from Sigma-Aldrich, St. Louis, Mo. (www.sigma-aldrich.com).

Compounds of general formula R¹ (CH₂)_(n)—X wherein R¹ is

are obtainable by reacting compounds of general formula R (CH₂)_(n)—Xwherein R¹ is —CN, X is a halogen, and n is an integer ranging from 1 to4 with TMSN₃ in the presence of tin oxide as described in S. J.Wittenberg et al., J. Org. Chem. 58:4139-4141 (1993).

Certain Triazaspiro Compounds can have asymmetric centers and thereforeexist in different enantiomeric and diastereomeric forms. A TriazaspiroCompound can be in the form of an optical isomer or a diastereomer.Accordingly, the invention encompasses Triazaspiro Compounds and theiruses as described herein in the form of their optical isomers,diasteriomers, and mixtures thereof, including a racemic mixture.

In addition, one or more hydrogen, carbon or other atoms of aTriazaspiro Compound can be replaced by an isotope of the hydrogen,carbon or other atoms. Such compounds, which are encompassed by thepresent invention, are useful as research and diagnostic tools inmetabolism pharmacokinetic studies and in binding assays.

4.5 Therapeutic Uses of the Triazaspiro Compounds

In accordance with the invention, the Triazaspiro Compounds areadministered in certain embodiments, to an animal, e.g. a mammal or ahuman, for the treatment or prevention of pain. The TriazaspiroCompounds can be used to treat or prevent acute or chronic pain. Forexample, the Triazaspiro Compounds can be used for, but are not limitedto, treating or preventing cancer pain, central pain, labor pain,myocardial infarction pain, pancreatic pain, colic pain, post-operativepain, headache pain, muscle pain, and pain associated with intensivecare.

The Triazaspiro Compounds can also be used for inhibiting, preventing,or treating pain associated with inflammation or with an inflammatorydisease in an animal. The pain to be inhibited, treated or prevented maybe associated with inflammation associated with an inflammatory disease,which can arise where there is an inflammation of the body tissue, andwhich can be a local inflammatory response and/or a systemicinflammation. For example, the Triazaspiro Compounds can be used toinhibit, treat, or prevent pain associated with inflammatory diseasesincluding, but not limited to: organ transplant rejection; reoxygenationinjury resulting from organ transplantation (see Grupp et al,. J. Mol.Cell Cardiol. 31:297-303 (1999)) including, but not limited to,transplantation of the heart, lung, liver, or kidney; chronicinflammatory diseases of the joints, including arthritis, rheumatoidarthritis, osteoarthritis and bone diseases associated with increasedbone resorption; inflammatory bowel diseases, such as ileitis,ulcerative colitis, Barrett's syndrome, and Crohn's disease;inflammatory lung diseases, such as asthma, adult respiratory distresssyndrome, and chronic obstructive airway disease; inflammatory diseasesof the eye, including corneal dystrophy, trachoma, onchocerciasis,uveitis, sympathetic ophthalmitis and endophthalmitis; chronicinflammatory diseases of the gum, including gingivitis andperiodontitis; tuberculosis; leprosy; inflammatory diseases of thekidney, including uremic complications, glomerulonephritis andnephrosis; inflammatory diseases of the skin, includingsclerodermatitis, psoriasis and eczema; inflammatory diseases of thecentral nervous system, including chronic demyclinating diseases of thenervous system, multiple sclerosis, AIDS-related neurodegeneration andAlzheimer s disease, infectious meningitis, encephalomyelitis,Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosisand viral or autoimmune encephalitis; autoimmune diseases, includingType I and Type II diabetes mellitus; diabetic complications, including,but not limited to, diabetic cataract, glaucoma, retinopathy,nephropathy (such as microaluminuria and progressive diabeticnephropathy), polyneuropathy, mononeuropathies, autonomic neuropathy,gangrene of the feet, atherosclerotic coronary arterial disease,peripheral arterial disease, nonketotic hyperglycemic-hyperosmolar coma,foot ulcers, joint problems, and a skin or mucous membrane complication(such as an infection, a shin spot, a candidal infection or necrobiosislipoidica diabeticorum); immune-complex vasculitis, and systemic lupuserythematosus (SLE); inflammatory diseases of the heart, such ascardiomyopathy, ischemic heart disease hypercholesterolemia, andatherosclerosis; as well as various other diseases that can havesignificant inflammatory components, including preeclampsia, chronicliver failure, brain and spinal cord trauma, and cancer. The TriazaspiroCompounds can also be used for inhibiting, treating, or preventing painassociated with inflammatory disease that can, for example, be asystemic inflammation of the body, exemplified by gram-positive or gramnegative shock, hemorrhagic or anaphylactic shock, or shock induced bycancer chemotherapy in response to pro-inflammatory cytokines, e.g.,shock associated with pro-inflammatory cytokines. Such shock can beinduced, e.g., by a chemotherapeutic agent that is administered as atreatment for cancer.

Without wishing to be bound by theory, it is believed that theTriazaspiro Compounds are agonists for an opioid receptor.

The invention also relates to methods for stimulating opioid-receptorfunction in a cell comprising contacting a cell capable of expressing anopioid receptor with an effective amount of a Triazaspiro Compound. Themethod is also useful for stimulating opioid receptor function in a cellin vivo, in an animal, such as a human, by contacting a cell capable ofexpressing an opioid receptor, in an animal, with an effective amount ofa Triazaspiro Compound. In one embodiment, the method is useful fortreating or preventing pain in an animal. Brain tissue, spinal chordtissue, immune cells, cells of the gastrointestinal tract, and primaryafferent nerve cells are examples of tissues and/or cells that canexpress an opioid receptor. This method can be used in vitro, forexample, as an assay to select cells that express an opioid receptor.

4.5.1 Therapeutic/Prophylactic Administration and Compositions of theInvention

Due to their activity, the Triazaspiro Compounds are advantageouslyuseful in veterinary and human medicine. As described above, theTriazaspiro Compounds are useful for treating or preventing pain in ananimal in need thereof.

When administered to an animal, the Triazaspiro Compounds can beadministered as a component of a composition that comprises apharmaceutically acceptable carrier or excipient. The presentcompositions, which comprise a Triazaspiro Compound, are, in certainembodiments, administered orally. The compositions of the invention canalso be administered by any other convenient route, for example, byinfusion or bolus injection, by absorption through epithelial ormucocutaneous linings (e.g., oral mucosa, rectal, and intestinal mucosa,etc.) and can be administered together with another therapeutic agent.Administration can be systemic or local. Various delivery systems areknown, e.g., encapsulation in liposomes, microparticles, microcapsules,capsules, etc., and can be used to administer the Triazaspiro Compounds.

Methods of administration include but are not limited to intradermal,intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal,epidural, oral, sublingual, intracerebral, intravaginal, transdermal,rectal, by inhalation, or topical, particularly to the ears, nose, eyes,or skin. The mode of administration is left to the discretion of thepractitioner. In most instances, administration will result in therelease of the Triazaspiro Compounds into the bloodstream.

In specific embodiments, the Triazaspiro Compounds can be administeredlocally. This can be achieved, for example, and not by way oflimitation, by local infusion during surgery, topical application, e.g.,in conjunction with a wound dressing after surgery, by injection, bymeans of a catheter, by means of a suppository, or by means of animplant, said implant being of a porous, non-porous, or gelatinousmaterial, including membranes, such as sialastic membranes, or fibers.

Pulmonary administration can also be employed, e.g., by use of aninhaler or nebulizer, and formulation with an aerosolizing agent, or viaperfusion in a fluorocarbon or synthetic pulmonary surfactant. Incertain embodiments, the Triazaspiro Compounds can be formulated as asuppository, with traditional binders and excipients such astriglycerides.

In another embodiment, the Triazaspiro Compounds can be delivered in avesicle, in particular a liposome (see Langer, Science 249:1527-1533(1990) and Treat et al., Liposomes in the Therapy of Infectious Diseaseand Cancer 317-327 and 353-365 (1989).

In yet another embodiment, the Triazaspiro Compounds can be delivered ina controlled-release system (see, e.g., Goodson, in Medical Applicationsof Controlled Release, supra, vol. 2, pp. 115-138 (1984)). Othercontrolled-release systems discussed in the review by Langer, Science249:1527-1533 (1990) may be used. In one embodiment, a pump may be used(Langer, Science 249:1527-1533 (1990); Sefton, CRC Crit. Ref. Biomed.Eng. 14:201 (1987); Buchwald et al., Surgery 88:507 (1980); and Saudeket al., N. Engl. J. Med. 321:574 (1989)). In another embodiment,polymeric materials can be used (see Medical Applications of ControlledRelease (Langer and Wise eds., 1974); Controlled Drug Bioavailability,Drug Product Design and Performance (Smolen and Ball eds., 1984); Rangerand Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:61 (1983); Levy etal., Science 228:190 (1985); During et al., Ann. Neurol. 25:351 (1989);and Howard et al., J. Neurosurg. 71:105 (1989)). In yet anotherembodiment, a controlled-release system can be placed in proximity of atarget of the Triazaspiro Compound thus requiring only a fraction of thesystemic dose.

The present compositions can optionally comprise a suitable amount of apharmaceutically acceptable excipient so as to provide the form forproper administration to the animal.

Such pharmaceutical excipients can be liquids, such as water and oils,including those of petroleum, animal, vegetable, or synthetic origin,such as peanut oil, soybean oil, mineral oil, sesame oil and the like.The pharmaceutical excipients can be saline, gum acacia, gelatin, starchpaste, talc, keratin, colloidal silica, urea, and the like. In addition,auxiliary, stabilizing, thickening, lubricating, and coloring agents maybe used. When administered to an animal, the pharmaceutically acceptableexcipients are, in certain embodiments, sterile. Water is a particularlyuseful excipient when the Triazaspiro Compound is administeredintravenously. Saline solutions and aqueous dextrose and glycerolsolutions can also be employed as liquid excipients, particularly forinjectable solutions. Suitable pharmaceutical excipients also includestarch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk,silica gel, sodium stearate, glycerol monostearate, talc, sodiumchloride, dried skim milk, glycerol, propylene, glycol, water, ethanol,and the like. The present compositions, if desired, can also containminor amounts of wetting or emulsifying agents, or pH buffering agents.

Suitable pharmaceutically acceptable carriers or excipients forintravenous administration of the Triazaspiro Compounds include, but arenot limited to, normal (about 0.9%) saline, about 25 to about 30%polyethylene glycol (“PEG”) diluted with saline or water, and about 2 toabout 30% hydroxy propyl β-cyclodextrin diluted with water.

Suitable pharmaceutically acceptable carriers or excipients forintraperitoneal administration of the Triazaspiro Compounds include, butare not limited to, normal (about 0.9%) saline, about 25 to about 30%polyethylene glycol (“PEG”) diluted with saline or water, about 25 toabout 30% propylene glycol (PG) diluted with saline or water, and about2 to about 30% hydroxypropyl β-cyclodextrin diluted with water.

Suitable pharmaceutically acceptable carriers or excipients forsubcutaneous and intramuscular administration of the TriazaspiroCompounds include, but are not limited to, normal (about 0.9%) saline,about 25 to about 30% polyethylene glycol (“PEG”) diluted with saline orwater, about 25 to about 30% propylene glycol (PG) diluted with salineor water, and water.

Suitable pharmnaceutically acceptable carriers or excipients for oraladministration of the Triazaspiro Compounds include, but are not limitedto, normal (about 0.9%) saline, about 25 to about 30% polyethyleneglycol (“PEG”) diluted with saline or water, about 2 to about 30%hydroxypropyl β-cyclodextrin diluted with water, about 25 to about 30%propylene glycol (PG) diluted with saline or water, about 1 to about 5%methylcellulose diluted with water, and water.

Suitable pharmaceutically acceptable carriers or excipients forintracerebroventricular and intrathecal administration of theTriazaspiro Compounds include, but are not limited to, normal (about0.9%) saline.

The present compositions can take the form of solutions, suspensions,emulsion, tablets, pills, pellets, capsules, capsules containingliquids, powders, sustained-release formulations, suppositories,aerosols, sprays, suspensions, or any other form suitable for use. Inone embodiment, the composition is in the form of a capsule (see e.g.,U.S. Pat. No. 5,698,155). Other examples of suitable pharmaceuticalexcipients are described in Remington 's Pharmaceutical Sciences1447-1676 (Alfonso R. Gennaro ed., 19th ed. 1995), incorporated hereinby reference.

In one embodiment, the Triazaspiro Compounds are formulated inaccordance with routine procedures as a composition adapted for oraladministration to an animal, particularly a human being. Compositionsfor oral delivery can be in the form of tablets, lozenges, aqueous oroily suspensions, granules, powders, emulsions, capsules, syrups, orelixirs, for example. Orally administered compositions can contain oneor more agents, for example, sweetening agents such as fructose,aspartame or saccharin; flavoring agents such as peppermint, oil ofwintergreen, or cherry; coloring agents; and preserving agents, toprovide a pharmaceutically palatable preparation. Moreover, where intablet or pill form, the compositions can be coated to delaydisintegration and absorption in the gastrointestinal tract therebyproviding a sustained action over an extended period of time.Selectively permeable membranes surrounding an osmotically activedriving compound are also suitable for orally administered compositions.In these later platforms, fluid from the environment surrounding thecapsule is imbibed by the driving compound, which swells to displace theagent or agent composition through an aperture. These delivery platformscan provide an essentially zero-order delivery profile as opposed to thespiked profiles of immediate-release formulations. A time delay materialsuch as glycerol monostearate or glycerol stearate can also be used.Oral compositions can include standard excipients such as mannitol,lactose, starch, magnesium stearate, sodium saccharin, cellulose andmagnesium carbonate. Such excipients are, in certain embodiments, ofpharmaceutical grade.

In another embodiment, the Triazaspiro Compounds can be formulated forintravenous administration. Typically, compositions for intravenousadministration comprise sterile isotonic aqueous buffer. Wherenecessary, the compositions can also include a solubilizing agent.Compositions for intravenous administration can optionally include alocal anesthetic such as lignocaine to lessen any pain at the site ofthe injection. Generally, the ingredients are supplied either separatelyor mixed together in unit dosage form, for example, as a dry lyophilizedpowder or water-free concentrate in a hermetically sealed container suchas an ampoule or sachette indicating the quantity of active agent. Wherethe Triazaspiro Compounds are to be administered by infusion, they canbe dispensed, for example, from an infusion bottle containing sterilepharmaceutical grade water or saline. Where the Triazaspiro Compoundsare administered by injection, an ampoule of sterile water for injectionor saline can be provided so that the ingredients can be mixed prior toadministration.

The Triazaspiro Compounds can be administered by controlled-releasemeans or by delivery devices that are well known to those of ordinaryskill in the art. Examples include, but are not limited to, thosedescribed in U.S. Pat. Nos. 3,845,770; 3,916,899; 3,536,809; 3,598,123;4,008,719; 5,674,533; 5,059,595; 5,591,767; 5,120,548; 5,073,543;5,639,476; 5,354,556; and 5,733,566, each of which is incorporatedherein by reference. Such dosage forms can be used to provide slow orcontrolled-release of one or more active ingredients using, for example,hydropropylmethyl cellulose, other polymer matrices, gels, permeablemembranes, osmotic systems, multilayer coatings, microparticles,liposomes, microspheres, or a combination thereof to provide the desiredrelease profile in varying proportions. Suitable controlled-releaseformulations known to those of ordinary skill in the art, includingthose described herein, can be readily selected for use with the activeingredients of the invention. The invention thus encompasses single unitdosage forms suitable for oral administration such as, but not limitedto, tablets, capsules, gelcaps, and caplets that are adapted forcontrolled-release.

Controlled-release pharmaceutical compositions can have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. In one embodiment a controlled-release compositioncomprises a minimal amount of a Triazaspiro Compound to cure or controlthe condition in a minimum amount of time. Advantages ofcontrolled-release compositions include extended activity of the drug,reduced dosage frequency, and increased patient compliance. In addition,controlled-release compositions can favorably affect the time of onsetof action or other characteristics, such as blood levels of theTriazaspiro Compound, and can thus reduce the occurrence of side (e.g.,adverse) effects.

Controlled-release compositions can initially release an amount of aTriazaspiro Compound that promptly produces the desired therapeuticeffect, and gradually and continually release other amounts of theTriazaspiro Compound to maintain this level of therapeutic orprophylactic effect over an extended period of time. To maintain thisconstant level of the Triazaspiro Compound in the body, the TriazaspiroCompound can be released from the dosage form at a rate that willreplace the amount of Triazaspiro Compound being metabolized andexcreted from the body. Controlled-release of an active ingredient canbe stimulated by various conditions including, but not limited to,changes in pH, changes in temperature, concentration or availability ofenzymes, concentration or availability of water, or other physiologicalconditions or compounds.

The amount of the Triazaspiro Compound that is effective in thetreatment or prevention of pain depends on the nature of the disorder orcondition causing the pain and can be determined by standard clinicaltechniques. In addition, in vitro or in vivo assays can optionally beemployed to help identify optimal dosage ranges. The precise dose to beemployed can also depend on the route of administration, and theseriousness of the pain and should be decided according to the judgmentof the practitioner and each patient's circumstances in view ofpublished clinical studies. Suitable effective amounts, however, range,in certain embodiments, from about 10 micrograms to about 2500milligrams about every 4 h, although typically about 100 mg or less. Incertain embodiments, the effective amount ranges from about 0.01milligrams to about 100 milligrams of a Triazaspiro Compound about every4 h, from about 0.020 milligrams to about 50 milligrams about every 4 h,and from about 0.025 milligrams to about 20 milligrams about every 4 h.The effective amounts described herein refer to total amountsadministered; that is, if more than one Triazaspiro Compound isadministered, the dosages correspond to the total amount administered.

Where a cell capable of expressing an opioid receptor is contacted witha Triazaspiro Compound in vitro, the effective amount will typicallyrange, in certain embodiments, from about 0.01 mg to about 100 mg/L,from about 0.1 mg to about 50 mg/L, and from about 1 mg to about 20mg/L, of a solution or suspension of a pharmaceutically acceptablecarrier or excipient.

Where a cell capable of expressing an opioid receptor is contacted witha Triazaspiro Compound in vivo, the effective amount will typicallyrange, in certain embodiments, from about 0.01 mg to about 100 mg/kg ofbody weight per day, from about 0.1 mg to about 50 mg/kg body weight perday, and from about 1 mg to about 20 mg/kg of body weight per day.

The Triazaspiro Compounds are, in certain embodiments, assayed in vitroor in vivo for the desired therapeutic or prophylactic activity prior touse in humans. Animal model systems can be used to demonstrate safetyand efficacy.

The present methods for treating or preventing pain in an animal canfurther comprise administering to the animal being administered aTriazaspiro Compound an effective amount of another therapeutic agent.

The present methods for stimulating opioid-receptor function in a cellcan further comprise contacting the cell with an effective amount ofanother therapeutic agent.

Examples of other therapeutic agents include, but are not limited to, anopioid agonist, a non-opioid analgesic, a non-steroid anti-inflammatoryagent, an antimigraine agent, a Cox-II inhibitor, an antiemetic, aβ-adrenergic blocker, an anticonvulsant, an antidepressant, aCa2+-channel blocker, an anticancer agent, an anti-anxiety agent, anagent for treating or preventing an addictive disorder and mixturesthereof.

Effective amounts of the other therapeutic agents are well known tothose skilled in the art. However, it is well within the skilledartisan's purview to determine the other therapeutic agent's optimaleffective-amount range. In one embodiment of the invention where anothertherapeutic agent is administered to an animal, the effective amount ofthe Triazaspiro Compound is less than its effective amount would bewhere the other therapeutic agent is not administered. In this case,without being bound by theory, it is believed that the TriazaspiroCompound and the other therapeutic agent act synergistically to treat orprevent pain.

Examples of useful opioid agonists include, but are not limited to,alfentanil, allylprodine, alphaprodine, anileridine, benzylmorphine,bezitramide, buprenorphine, butorphanol, clonitazene, codeine,desomorphine, dextromoramide, dezocine, diampromide, diamorphone,dihydrocodeine, dihydromorphine, dimenoxadol, dimepheptanol,dimethylthiambutene, dioxaphetyl butyrate, dipipanone, eptazocine,ethoheptazine, ethylmethylthiambutene, ethylmorphine, etonitazenefentanyl, heroin, hydrocodone, hydromorphone, hydroxypethidine,isomethadone, ketobemidone, levorphanol, levophenacylmorphan,lofentanil, meperidine, meptazinol, metazocine, methadone, metopon,morphine, myrophine, nalbuphine, narceine, nicomorphine, norlevorphanol,normethadone, nalorphine, normorphine, norpipanone, opium, oxycodone,oxymorphone, papaveretum, pentazocine, phenadoxone, phenomorphan,phenazocine, phenoperidine, piminodine, piritramide, proheptazine,promedol, properidine, propiram, propoxyphene, sufentanil, tilidine,tramadol, pharmaceutically acceptable salts thereof, and mixturesthereof.

In certain embodiments, the opioid agonist is selected from codeine,hydromorphone, hydrocodone, oxycodone, dihydrocodeine, dihydromorphine,morphine, tramadol, oxymorphone, pharmaceutically acceptable saltsthereof, and mixtures thereof.

Examples of useful non-opioid analgesics include non-steroidalanti-inflammatory agents, such as aspirin, ibuprofen, diclofenac,naproxen, benoxaprofen, flurbiprofen, fenoprofen, flubufen, ketoprofen,indoprofen, piroprofen, carprofen, oxaprozin, pramoprofen, muroprofen,trioxaprofen, suprofen, aminoprofen, tiaprofenic acid, fluprofen,bucloxic acid, indomethacin, sulindac, tolmetin, zomepirac, tiopinac,zidometacin, acemetacin, fentiazac, clidanac, oxpinac, mefenamic acid,meclofenamic acid, flufenamic acid, niflumic acid, tolfenamic acid,diflurisal, flufenisal, piroxicam, sudoxicam, isoxicam, andpharmaceutically acceptable salts thereof, and mixtures thereof.Examples of other suitable non-opioid analgesics include the following,non-limiting, chemical classes of analgesic, antipyretic, nonsteroidalantiinflammatory drugs: salicylic acid derivatives, including aspirin,sodium salicylate, choline magnesium trisalicylate, salsalate,diflunisal, salicylsalicylic acid, sulfasalazine, and olsalazin;para-aminophennol derivatives including acetaminophen and phenacetin;indole and indene acetic acids, including indomethacin, sulindac, andetodolac; heteroaryl acetic acids, including tolmetin, diclofenac, andketorolac; anthranilic acids (fenamates), including mefenamic acid, andmeclofenamic acid; enolic acids, including oxicams (piroxicam,tenoxicam), and pyrazolidinediones (phenylbutazone, oxyphenthartazone);and alkanones, including nabumetone. For a more detailed description ofthe NSAIDs, see Paul A. Insel, Analgesic-Antipyretic andAntiinflammatory Agents and Drugs Employed in the Treatment of Gout, inGoodman & Gilman's The Pharmacological Basis of Therapeutics 617-57(Perry B. Molinhoff and Raymond W. Ruddon eds., 9^(th) ed 1996) and GlenR. Hanson, Analgesic, Antipyretic and Anti-Inflammatory Drugs inRemington: The Science and Practice of Pharmacy Vol II 1196-1221 (A. R.Gennaro ed. 19th ed. 1995) which are hereby incorporated by reference intheir entireties. Suitable Cox-II inhibitors and 5-lipoxygenaseinhibitors, as well as combinations thereof, are described in U.S. Pat.No. 6,136,839, which is hereby incorporated by reference in itsentirety. Cox-II inhibitors include, but are not limited to, rofecoxiband celecoxib.

Examples of useful antimigraine agents include, but are not limited to,alpiropride, dihydroergotamine, dolasetron, ergocomine, ergocominine,ergocryptine, ergot, ergotamine, flumedroxone acetate, fonazine,lisuride, lomerizine, methysergide oxetorone, pizotyline, and mixturesthereof.

The other therapeutic agent can also be an agent useful for reduce anypotential side effects of a Triazaspiro Compound. For example, the othertherapeutic agent can be an antiemetic agent. Useful antiemetic agentsinclude, but are not limited to, metoclopromide, domperidone,prochlorperazine, promethazine, chlorpromazine, trimethobenzamide,ondansetron, granisetron, hydroxyzine, acetylleucine monoethanolamine,alizapride, azasetron, benzquinamide, bietanautine, bromopride,buclizine, clebopride, cyclizine, dimenhydrinate, diphenidol,dolasetron, meclizine, methallatal, metopimazine, nabilone, oxypemdyl,pipamazine, scopolamine, sulpiride, tetrahydrocannabinol,thiethylperazine, thioproperazine, tropisetron, and mixtures thereof.

Examples of useful β-adrenergic blockers include, but are not limitedto, acebutolol, alprenolol, amosulabol, arotinolol, atenolol, befunolol,betaxolol, bevantolol, bisoprolol, bopindolol, bucumolol, bufetolol,bufuralol, bunitrolol, bupranolol, butidrine hydrochloride, butofilolol,carazolol, carteolol, carvedilol, celiprolol, cetamolol, cloranolol,dilevalol, epanolol, esmolol, indenolol, labetalol, levobunolol,mepindolol, metipranolol, metoprolol, moprolol, nadolol, nadoxolol,nebivalol, nifenalol, nipradilol, oxprenolol, penbutolol, pindolol,practolol, pronethalol, propranolol, sotalol, sulfinalol, talinolol,tertatolol, tilisolol, timolol, toliprolol, and xibenolol.

Examples of useful anticonvulsants include, but are not limited to,acetylpheneturide, albutoin, aloxidone, aminoglutethimide,4-amino-3-hydroxybutyric acid, atrolactamide, beclamide, buramate,calcium bromide, carbamazepine, cinromide, clomethiazole, clonazepam,decimemide, diethadione, dimethadione, doxenitroin, eterobarb,ethadione, ethosuximide, ethotoin, felbamate, fluoresone, gabapentin,5-hydroxytryptophan, lamotrigine, magnesium bromide, magnesium sulfate,mephenytoin, mephobarbital, metharbital, methetoin, methsuximide,5-methyl-5-(3-phenanthryl)-hydantoin, 3-methyl-5-phenylhydantoin,narcobarbital, nimetazepam, nitrazepam, oxcarbazepine, paramethadione,phenacemide, phenetharbital, pheneturide, phenobarbital, phensuximide,phenylmethylbarbituric acid, phenytoin, phethenylate sodium, potassiumbromide, pregabaline, primidone, progabide, sodium bromide, solanum,strontium bromide, suclofenide, sulthiame, tetrantoin, tiagabine,topiramate, trimethadione, valproic acid, valpromide, vigabatrin, andzonisamide.

Examples of useful antidepressants include, but are not limited to,binedaline, caroxazone, citalopram, dimethazan, fencamine, indalpine,indeloxazine hydrocholoride, nefopam, nomifensine, oxitriptan,oxypertine, paroxetine, sertraline, thiazesim, trazodone, benmoxine,iproclozide, iproniazid, isocarboxazid, nialamide, octamoxin,phenelzine, cotinine, rolicyprine, rolipram, maprotiline, metralindole,mianserin, mirtazepine, adinazolam, amitriptyline, amitriptylinoxide,amoxapine, butriptyline, clomipramine, demexiptiline, desipramine,dibenzepin, dimetacrine, dotbiepin, doxepin, fluacizine, imipramine,imipramine N-oxide, iprindole, lofepramine, melitracen, metapramine,nortriptyline, noxiptilin, opipramol, pizotyline, propizepine,protriptyline, quinupramine, tianeptine, trimipramine, adrafinil,benactyzine, bupropion, butacetin, dioxadrol, duloxetine, etoperidone,febarbamate, femoxetine, fenpentadiol, fluoxetine, fluvoxamine,hematoporphyrin, hypericin, levophacetoperane, medifoxamine,milnacipran, minaprine, moclobemide, nefazodone, oxaflozane, piberaline,prolintane, pyrisuccideanol, ritanserin, roxindole, rubidium chloride,sulpiride, tandospirone, thozalinone, tofenacin, toloxatone,tranylcypromine, L-tryptophan, venlafaxine, viloxazine, and zimeldine.

Examples of useful Ca2+-channel blockers include, but are not limitedto, bepridil, clentiazem, diltiazem, fendiline, gallopamil, mibefradil,prenylamine, semotiadil, terodiline, verapamil, amlodipine, aranidipine,barnidipine, benidipine, cilnidipine, efonidipine, elgodipine,felodipine, isradipine, lacidipine, lercanidipine, manidipine,nicardipine, nifedipine, nilvadipine, nimodipine, nisoldipine,nitrendipine, cinnarizine, flunarizine, lidoflazine, lomerizine,bencyclane, etafenone, fantofarone, and perhexiline.

Examples of useful anticancer agents include, but are not limited to,acivicin, aclarubicin, acodazole hydrochloride, acronine, adozelesin,aldesleukin, altretamine, ambomycin, ametantrone acetate,aminoglutethimide, amsacrine, anastrozole, anthramycin, asparaginase,asperlin, azacitidine, azetepa, azotomycin, batimastat, benzodepa,bicalutamide, bisantrene hydrochloride, bisnafide dimesylate, bizelesin,bleomycin sulfate, brequinar sodium, bropirimine, busulfan,cactinomycin, calusterone, caracemide, carbetimer, carboplatin,carmustine, carubicin hydrochloride, carzelesin, cedefingol,chlorambucil, cirolemycin, cisplatin, cladribine, crisnatol mesylate,cyclophosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicinhydrochloride, decitabine, dexormaplatin, dezaguanine, dezaguaninemesylate, diaziquone, docetaxel, doxorubicin, doxorubicin hydrochloride,droloxifene, droloxifene citrate, dromostanolone propionate, duazomycin,edatrexate, eflomithine hydrochloride, elsamitrucin, enloplatin,enpromate, epipropidine, epirubicin hydrochloride, erbulozole,esorubicin hydrochloride, estramustine, estramustine phosphate sodium,etanidazole, etoposide, etoposide phosphate, etoprine, fadrozolehydrochloride, fazarabine, fenretinide, floxuridine, fludarabinephosphate, fluorouracil, flurocitabine, fosquidone, fostriecin sodium,gemcitabine, gemcitabine hydrochloride, hydroxyurea, idarubicinhydrochloride, ifosfamide, ilmofosine, interleukin II (includingrecombinant interleukin II or rIL2), interferon alfa-2a, interferonalfa-2b, interferon alfa-n1, interferon alfa-n3, interferon beta-I a,interferon gamma-I b, iproplatin, irinotecan hydrochloride, lanreotideacetate, letrozole, leuprolide acetate, liarozole hydrochloride,lometrexol sodium, lomustine, losoxantrone hydrochloride, masoprocol,maytansine, mechlorethamine hydrochloride, megestrol acetate,melengestrol acetate, melphalan, menogaril, mercaptopurine,methotrexate, methotrexate sodium, metoprine, meturedepa, mitindomide,mitocarcin, mitocromin, mitogillin, mitomalcin, mitomycin, mitosper,mitotane, mitoxantrone hydrochloride, mycophenolic acid, nocodazole,nogalamycin, ormnaplatin, oxisuran, paclitaxel, pegaspargase,peliomycin, pentamustine, peplomycin sulfate, perfosfamide, pipobroman,piposulfan, piroxantrone hydrochloride, plicamycin, plomestane, porfimersodium, porfiromycin, prednimustine, procarbazine hydrochloride,puromycin, puromycin hydrochloride, pyrazofurin, riboprine, rogletimide,safingol, safingol hydrochloride, semustine, simtrazene, sparfosatesodium, sparsomycin, spirogermanium hydrochloride, spiromustine,spiroplatin, streptonigrin, streptozocin, sulofenur, talisomycin,tecogalan sodium, tegafur, teloxantrone hydrochloride, temoporfin,teniposide, teroxirone, testolactone, thiamiprine, thioguanine,thiotepa, tiazofurin, tirapazamine, toremifene citrate, trestoloneacetate, triciribine phosphate, trimetrexate, trimetrexate glucuronate,triptorelin, tubulozole hydrochloride, uracil mustard, uredepa,vapreotide, verteporfin, vinblastine sulfate, vincristine sulfate,vindesine, vindesine sulfate, vinepidine sulfate, vinglycinate sulfate,vinleurosine sulfate, vinorelbine tartrate, vinrosidine sulfate,vinzolidine sulfate, vorozole, zeniplatin, zinostatin, zorubicinhydrochloride.

Examples of other useful anti-cancer agents include, but are not limitedto, 20-epi-1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone;aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TKantagonists; altretamine; ambamustine; amidox; amifostine;aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole;andrographolide; angiogenesis inhibitors; antagonist D; antagonist G;antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen,prostatic carcinoma; antiestrogen; antineoplaston; antisenseoligonucleotides; aphidicolin glycinate; apoptosis gene modulators;apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; argininedeaminase; asulacrine; atamestane; atrimustine; axinastatin 1;axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatinIII derivatives; balanol; batimastat; BCR/ABL antagonists;benzochlorins; benzoylstaurosporine; beta lactam derivatives;beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor;bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistrateneA; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine;calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2;capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRestM3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinaseinhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorlns;chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine;clomifene analogues; clotrimazole; collismycin A; collismycin B;combretastatin A4; combretastatin analogue; conagenin; crambescidin 816;crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A;cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate;cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B;deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;diaziquone; didemnin B; didox; diethylnorspermine;dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin; diphenylspiromustine; docetaxel; docosanol; dolasetron; doxifluridine;droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine;edelfosine; edrecolomab; eflomithine; elemene; emitefur; epirubicin;epristeride; estramustine analogue; estrogen agonists; estrogenantagonists; etanidazole; etoposide phosphate; exemestane; fadrozole;fazarabine; fenretinide; filgrastim; finasteride; flavopiridol;flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras famesyl protein transferase inhibitors; rasinhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B 1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer.

Therapeutic agents useful for treating or preventing an addictivedisorder include, but are not limited to, methadone, desipramine,amantadine, fluoxetine, buprenorphine, an opiate agonist,3-phenoxypyridine, or a serotonin antagonist.

Examples of useful anti-anxiety agents include, but are not limited to,benzodiazepines, such as alprazolam, chlordiazepoxide, clonazepam,clorazepate, diazepam, halazepam, lorazepam, oxazepam, and prazepam;non-benzodiazepine agents, such as buspirone; and tranquilizers, such asbarbituates.

A Triazaspiro Compound and the other therapeutic agent can actadditively or, in one embodiment, synergistically. In one embodiment, aTriazaspiro Compound is administered concurrently with anothertherapeutic agent. In one embodiment, a composition comprising aneffective amount of a Triazaspiro Compound and an effective amount ofanother therapeutic agent can be administered. Alternatively, acomposition comprising an effective amount of a Triazaspiro Compound anda different composition comprising an effective amount of anothertherapeutic agent can be concurrently administered. In anotherembodiment, an effective amount of a Triazaspiro Compound isadministered prior or subsequent to administration of an effectiveamount of another therapeutic agent. In this embodiment, the TriazaspiroCompound is administered while the other therapeutic agent exerts itstherapeutic effect, or the other thereapeutic agent is administeredwhile the Triazaspiro Compound exerts its preventive or therapeuticeffect for treating or preventing pain.

In another embodiment, a composition of the invention is prepared by amethod comprising admixing a Triazaspiro Compound or pharmaceuticallyacceptable salt and a pharmaceutically acceptable carrier or excipient.Admixing can be accomplished using methods well known for admixing acompound (or salt) and a pharmaceutically acceptable carrier orexcipient. In one embodiment, the Triazaspiro Compound or thepharmaceutically acceptable salt of the Compound is present in thecomposition in an effective amount.

4.5.2 Kits

The invention encompasses kits that can simplify the administration of aTriazaspiro Compound to an animal.

A typical kit of the invention comprises a unit dosage form of aTriazaspiro Compound. In one embodiment, the unit dosage form is acontainer, which in certain embodiments, is a sterile container,containing an effective amount of a Triazaspiro Compound and apharmaceutically acceptable carrier or excipient. The kit can furthercomprise a label or printed instructions instructing the use of theTriazaspiro Compound to treat or prevent pain. The kit can also furthercomprise a unit dosage form of another therapeutic agent, for example, acontainer containing an effective amount of the other therapeutic agent.In one embodiment, the kit comprises a container containing an effectiveamount of a Triazaspiro Compound and an effective amount of anothertherapeutic agent. Examples of other therapeutic agents include, but arenot limited to, those listed above.

Kits of the invention can further comprise a device that is useful foradministering the unit dosage forms. Examples of such devices include,but are not limited to, syringes, drip bags, patches, and inhalers.

The following examples are set forth to assist in understanding theinvention and should not, of course, be construed as specificallylimiting the invention described and claimed herein. Such variations ofthe invention, including the substitution of all equivalents now knownor later developed, which would be within the purview of those skilledin the art, and changes in formulation or minor changes in experimentaldesign, are to be considered to fall within the scope of the inventionincorporated herein.

5. EXAMPLES

Examples 1-21 relate to the synthesis of illustrative TriazaspiroCompounds.

5.1 Example 1 Synthesis of Compound AC

To a mixture of Compound A (2 mmol) and Compound B (2 mmol) in 6 mL ofacetonitrile was added 1.5 eq. of diisopropylethylamine and theresulting mixture was heated to 60° C. and allowed to stir for about 12h. Thin layer chromatography indicated the complete disappearance ofCompound A. The solvent was removed under reduced pressure. Theresulting residue was dissolved in 10 mL of ethyl acetate, washed withwater (10 mL, 2 times), the organic layer dried (K₂CO₃), and the solventremoved under reduced pressure. The resulting residue was purified bycolumn chromatography using a silica column eluted with 5%triethylamine, 25% ethyl acetate, and 70% hexane to provide 120 mg ofCompound AC (yield 10.6%). Compound AC was shown to be greater than 97%pure by high pressure liquid chromatography (HPLC) analysis.

The identity of Compound AC was confirmed using ¹H NMR and mass spectral(MS) analysis.

¹H NMR (CDCl₃): δ 1.6 (m, 2H), 2.2 (m, 2H), 2.55-2.8 (m, 8H), 3.7 (s,3H), 4.75 (s, 2H), 6.5 (bs, 1H), 6.9 (m, 3H), 7.2-7.4 (m, 11H). MS: m/z484.2.

5.2 Example 2 Synthesis of Compound AF

Compound AF was prepared by dissolving Compound AC (0.5 mmol) in 3 mL ofmethanol and 40 mL of 40% aqueous potassium hydroxide and heating theresulting mixture at 80° C. with stirring for about 2 h. LC/MS indicateddisappearance of Compound AC. Then 2 mL of water and 2 mL of 3N HCl wasadded to the mixture to adjust the pH to a value of about 1. Theresulting precipitate was collected and dried under vacuum to provide110 mg of Compound AF. Compound AF was shown to be greater than 97% pureby HPLC analysis.

The identity of compound AF was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): δ 2.0 (d, 2H), 2.65-2.75 (m, 2H), 2.8-3.0 (m, 4H), 3.5(m, 2H), 2.7 (m, 2H), 4.7 (s, 2H), 6.9-7.05 (m, 2H), 7.2-7.4 (m, 13H).MS: m/z 470.2.

5.3 Example 3 Synthesis of Compound AA

To a mixture of Compound A (3.6 mmol) (commercially available fromSigma-Aldrich, St. Louis, Mo., www.sigma-aldrich.com) and Compound B(3)(3.6 mmole) (commercially available from Sigma-Aldrich, St. Louis, Mo.,www.sigma-aldrich.com) in 50 mL of acetonitrile was added in one portion1.2 molar eq. of diisopropylethyamine (DIEA) and 3 drops ofdimethylformamide (DMF). The resulting mixture was heated to 60° C. andallowed to stir for about 12 h. Thin-layer chromatography (TLC)indicated the disappearance of Compound A. The solvent was removed underreduced pressure and 50 mL of 1N NaOH and 50 mL of ethyl acetate wereadded to the resulting residue. The organic layer was separated and theaqueous layer washed with ethyl acetate (50 mL, 2 times). The organiclayers were combined, dried (K₂CO₃), and the solvent removed underreduced pressure to provide a yellow oil that was purified using asilica gel column chromatography eluted with 5% triethylamine, 25% ethylacetate, and 70% hexane. The relevant fractions were combined andconcentrated to provide Compound AA. Compound AA was shown to be greaterthan 97% pure by HPLC analysis.

The identity of Compound AA was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.7 (d, 2H), 2.55 (dm, 2H), 2.65 (m, 4H), 2.7-2.9 (m,4H), 3.7 (s, 2H), 6.25 (bs, 1H), 6.8-6.95 (m, 3H), 7.3-7.5 (m, 12H). MS:m/z 451.2.

5.4 Example 4 Synthesis of Compound AK

To a solution of Compound AA (0.5 mmol) and trimethylsilylazide (1 mmol,2 eq.) in 3 mL of toluene was added dibutyl tin oxide (0.1 eq.) and theresulting mixture was heated at 110° C. for about 24 h. LC/MS indicatedcompete disappearance of Compound AA. The solvent was removed underreduced pressure. The resulting residue was dissolved in CHCl₃ andpurified using column chromatography on a florisil column eluted with 2%NH₄OH, 15% methanol, and 85% methylene chloride to provide 6 mg ofCompound AK as a pale yellow solid. Compound AK was shown to be greaterthan 97% pure by HPLC analysis.

The identity of compound AK was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): δ 2.0 (d, 2H), 2.9-3.0 (m, 4H), 3.1 (m, 2H), 3.4-3.5 (m,2H), 3.6-3.7 (m, 2H), 4.75 (s, 2H), 6.9 (t, 1H), 7.0-7.1 (m, 5H),7.2-7.3 (m, 5H), 7.35 (t, 2H), 7.8 (s, 2H). MS: m/z 494.

5.5 Example 5 Synthesis of Compound AB

4-Bromo-2,2-diphenylbutyric acid (Compound B(1), 23 g, 72 mmol) wassuspended in 150 mL of chloroform, and 20 mL of thionyl chloride (270mmol) was added dropwise. After addition of the thionyl chloride, 0.2 mLof dimethylformamide was added and the resulting solution heated atreflux for about 4 h. The reaction mixture was then concentrated underreduced pressure to provide 4-bromo-2,2-diphenylbutyric chloride(Compound D(1)) as a pale yellow oil that was used in the following stepwithout further purification.

To 100 mL of saturated aqueous Na₂CO₃ was added 50 mL of a 2M solutionof dimethylamine in tetrahydrofuran. The resulting solution was cooledto 0° C. and a solution of Compound D(1), prepared as described above,dissolved in 100 mL of toluene was added dropwise. The resulting mixturewas allowed to stir for about 12 h. The organic and aqueous layers ofthe reaction mixture were separated and the aqueous layer was extractedwith 30 mL of toluene and then extracted 3 times with 100 mL ofchloroform and the organic extracts were combined. The combined organicextracts were washed with water (30 mL), dried (K₂CO₃), and the solventwas removed under reduced pressure to provide a residue that wascrystallized from methyl isobutyl ketone to provide 12 g (53% yield) ofdimethyl(tetrahydro-3,3-diphenyl-2-furylidene)ammonium bromide (CompoundE(1)).

To a mixture of Compound A (1 mmol) and Compound E(1) (1 mmol) in 4 mLof dimethylformamide was added 3 eq. of Na₂CO₃. The resulting mixturewas heated at 80° C. with stirring for about 1.5 h. Thin layerchromatography showed the disappearance of Compound A. The solvent wasremoved under reduced pressure and 5 mL of 1 N NaOH and 10 mL of ethylacetate were added to the residue. The organic layer was separated andthe aqueous layer was washed with ethyl acetate (5 mL, 2 time). Theorganic layers were combined, dried (K₂CO₃), and the solvent removedunder reduced pressure to provide a yellow oil that was purified bycolumn chromatography using a silica column eluted with 5%triethylamine, 25% ethyl acetate, and 70% hexane to provide 284 mg ofCompound AB as a solid (58% yield); Compound AB was shown to be greaterthan 97% pure by HPLC analysis.

The identity of compound AB was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.6 (d, 2H), 2.2 (m, 2H), 2.35 (m, 2H), 2.45-2.6 (m,5H), 2.7 (m, 4H), 3.0 (m, 3H), 4.7 (s, 2H), 6.2 (bs, 1H), 6.8-6.9 (m,3H), 7.3 (m, 4H), 7.35-7.5 (m, 8H). MS: m/z 497.2.

5.6 Example 6 Synthesis of Compound AR

To 480 mg of Compound AB (1 mmol) in 5 mL of dimethylformamide was addedabut 1.2 eq. of NaH that had been washed twice with tetrahydrofuran. Gasevolution occurred and, after 2 min., 1.1 eq. of ethylbromopropionate(commercially available from Sigma-Aldrich, St. Louis, Mo.,www.sigma-aldrich.com) was added. The resulting reaction mixture wasallowed to stir for about 12 h. LC/MS indicated the disappearance ofcompound AA. Water (10 mL) and ethyl acetate (10 mL) were then added tothe reaction mixture. The organic layer was separated, dried (K₂CO₃),and the solvent was removed under reduced pressure. The resultingproduct was purified by column chromatography using a silica gel columneluted with 5% triethylamine, 25% ethyl acetate, and 70% hexane toprovide 400 mg of Compound AR as an oil (71% yield). Compound AR wasshown to be greater than 97% pure by HPLC analysis.

The identity of Compound AR was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.3 (t, 3H), 1.5 (d, 2H), 2.2 (m, 2H), 2.3 (bs, 3H),2.4-2.6 (m, 4H), 2.7 (m, 2H), 2.7-2.8 (m, 4H), 3.0 (bs, 3H), 3.7 (m,2H), 4.2 (q, 2H), 4.7 (s, 2H), 6.9 (m, 3H), 7.3 (m, 4H), 7.4 (m, 4H),7.45 (m, 4H). MS: m/z 597.

5.7 Example 7 Synthesis of Compound AT

Compound AT was prepared by dissolving Compound AR (0.5 mmol) in 3 mL ofmethanol and 40 mL of 40% aqueous potassium hydroxide and heating theresulting mixture at 80° C. with stirring for about 2 h. LC/MS indicateddisappearance of Compound AT. After completion of the reaction, 2 mL ofwater and 2 mL of 3N HCl was added to the mixture to adjust the pH to avalue of about 1. The resulting precipitate was collected and driedunder vacuum to provide 110 mg of Compound AT. Compound AT was shown tobe greater than 97% pure by HPLC analysis.

The identity of Compound AT was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.6 (d, 2H), 2.3 (s, 3H), 2.6 (m, 2H), 2.7 (m, 4H),3.0 (s, 3H), 3.3 (m, 4H), 3.7 (m, 4H), 4.7 (s, 2H), 6.8 (m, 1H), 7.0 (m,2H), 7.2 (m, 2H), 7.3 (m, 10H), 11.5 (bs, 1H). MS: m/z 569.

5.8 Example 8 Synthesis of Compound AH

450 mg (1 mmol) of Compound AA were added to about 1.2 mmol of NaH (thathad been washed twice with THF) in 10 mL of DMF. Gas evolution occurredand, after 2 min., 1.1 eq. of ethyl iodoacetate acid (1.1 mmol)(commercially available from Sigma-Aldrich, St. Louis, Mo.,www.sigma-aldrich.com) was added. The resulting reaction mixture wasallowed to stir for about 12 h. LC/MS indicated the disappearance ofCompound AA. Water (10 mL) and ethyl acetate (10 mL) were added to thereaction mixture. The organic layer was separated, dried (K₂CO₃), andthe solvent was removed under reduced pressure. The resulting productwas purified by column chromatography using a silica gel column elutedwith 5% triethylamine, 25% ethyl acetate, and 70% hexane to provide 390mg of Compound AH as an oil (72.7% yield). Compound AH was shown to begreater than 94% pure by HPLC analysis.

The identity of Compound AH was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.3 (t, 3H), 1.7 (d, 2H), 2.5-2.65 (m, 6H), 2.75 (m,2H), 2.85 (m, 2H), 4.1 (s, 2H), 4.2 (m, 2H), 4.75 (s, 2H), 6.85-6.95 (m,3H), 7.2-7.4 (m, 12H). MS: m/z 537.

5.9 Example 9 Synthesis of Compound AD

A solution of Compound AH (0.5 mmol) in 3 mL of methanol and 1 mL of 40%aqueous KOH was stirred and heated at 80° C. for about 2 h. LC/MS showeddisappearance of Compound AH. After 2 h, 2 mL of water and 2 mL of 3NHCl were added to the mixture to adjust the pH to a value of about 1.The resulting precipitate was isolated by filtration and dried underhigh vacuum to provide 130 mg of Compound AD (51.2% yield). Compound ADwas shown to be greater than 97% pure by HPLC analysis.

The identity of Compound AD was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): 67 2.2 (d, 2H), 2.7 (m, 2H), 3.0 (m, 2H), 3.2 (m, 2H),3.6 (m, 2H), 3.8 (m, 2H), 4.2 (s, 2H), 7.0 (m, 1H), 7.1 (m, 2H), 7.3-7.5(m, 12H). MS: m/z 509.

5.10 Example 10 Synthesis of Compound AG

450 mg (1 mmol) of Compound F was added to about 1.2 mmol of NaH (thathad been washed twice with THF) in 10 mL of DMF. Gas evolution occurredand, after 2 min., 1.1 eq. of ethyl iodoacetate (1.1 mmol) (commerciallyavailable from Sigma-Aldrich, St. Louis, Mo., www.sigma-aldrich.com) wasadded. The resulting reaction mixture was allowed to stir for about 12h. LC/MS indicated the disappearance of compound F. Water (10 mL) andethyl acetate (10 mL) were added to the reaction mixture. The organiclayer was separated, dried (K₂CO₃), and the solvent was removed underreduced pressure. The resulting product was purified by columnchromatography using a silica gel column eluted with 5% triethylamine,25% ethyl acetate, and 70% hexane) to provide 381 mg of Compound AG asan oil (74.7% yield). Compound AG was shown to be greater than 97% pureby HPLC analysis.

The identity of Compound AG was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): δ 1.3 (t, 3H), 1.7 (d, 2H), 2.45-2.65 (m, 6H), 2.77 (m,2H), 2.85 (m, 2H), 4.15 (s, 2H), 4.2 (m, 2H), 4.75 (s, 2H), 6.80-6.95(m, 3H), 7.2-7.4 (m, 12H). MS: m/z 512.

5.11 Example 11 Synthesis of Compound AE

A solution of Compound AG (0.5 mmol) in 3 mL of methanol and 1 mL of 40%aqueous KOH was stirred and heated at 80° C. for about 2 h. LC/MS showeddisappearance of Compound AG. After 2 h, 2 mL of water and 2 mL of 3NHCl were added to the mixture to adjust the pH to a value of about 1.The resulting precipitate was isolated by filtration and dried underhigh vacuum to provide 127 mg of Compound AE (52.5% yield). Compound AEwas shown to be greater than 97% pure by HPLC analysis.

The identity of Compound AE was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): δ 2.1 (d, 2H), 5.5-5.7 (m, 4H), 3.1 (m, 2H), 3.5 (m,2H), 3.75 (m, 2H), 4.0 (m, 3H), 6.9 (m, 1H), 7.0 (m, 2H), 7.2 (m, 2H),7.3 (m, 10H). MS: m/z 484.

5.12 Example 12 Synthesis of Compound AM

Compound F (0.9 mmol) in 5 mL of DMF was added to a flask containing 1.5eq. of NaH that had been flushed with argon. 2-Bromoacetamide (1.2 eq.)(commercially available from Sigma-Aldrich, St. Louis, Mo.,www.sigma-aldrich.com) was added to the flask and the resulting mixturewas allowed to stir at room temperature for 30 min. The mixture was thenheated to 60° C. and allowed to stir for about 12 h. The reactionmixture was then cooled to room temperature, diluted with 30 mL of ethylacetate, and washed with water (10 mL, 2 times). The organic layer wasseparated, dried (MgSO₄), and the solvent removed under reduced pressureto provide a residue that was purified using a florisil column elutedwith a mixture of 2% saturated aqueous NH₄0H, 28% methanol and 70%methylene chloride to provide 15 mg of Compound AM as a pale yellowsolid. Compound AM was shown to be greater than 97% pure by HPLCanalysis.

The identity of Compound AM was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.7 (d, 2H), 2.25 (m, 2H), 2.4 (m, 2H), 2.5 (m, 2H)2.8 (m, 4H), 4.0 (t, 1H), 4.05 (s, 2H), 4.8 (s, 2H), 5.4 (bs, 1H), 6.0(bs, 1H), 6.95 (m, 3H), 7.2 (m, 2H), 7.3 (m, 10H). MS: m/z 483.3.

5.13 Example 13 Synthesis of Compound AL

Compound AL was synthesized according to the method of Example 10 exceptthat BrCH₂CH₂NHSO₂CH₃ was used in place of ethyl iodoacetate.BrCH₂CH₂NHSO₂CH₃ can be prepared by reacting from 2-bromoethylamine(commercially available from Sigma-Aldrich, St. Louis, Mo.,www.sigma-aldrich.com) with methanesulfonyl chloride in methylenechloride in the presence of triethylamine at 0° as shown in the schemebelow:

The purity of Compound AL was determined to be greater than 97% by HPLCanalysis.

The identity of Compound AL was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): δ 1.7 (m, 2H), 2.3 (m, 2H), 2.4 (m, 2H) 2.5-2.6 (m, 2H),2.7-2.8 (m, 4H), 3.0 (s, 3H), 3.4 (m, 2H), 3.6 (m, 2H), 4.0 (t, 1H), 4.8(s, 2H), 5.1 (bs, 1II), 6.9 (m, 3H), 7.2 (m, 2H), 7.3 (m, 10H). MS: m/z547.

5.14 Example 14 Synthesis of Compounds AN and AO

Compound AN and Compound AO were synthesized according to the methods ofExample 10 and Example 11 except that ethyl 3-iodopropionate and ethyl5-bromovalerate (each commercially available from Sigma-Aldrich, St.Louis, Mo., www.sigma-aldrich.com), respectively, were used in place ofethyl iodoacetate. The purity of Compound AN and Compound AO was eachdetermined to be greater than 97% by HPLC analysis.

The identity of Compound AN was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.3 (m, 3H), 2.4 (m, 2H), 2.7-2.9 (m, 5H), 3.1 (m,4H), 3.3 (m, 2H), 3.65 (m, 2H), 3.9 (t, 1H), 4.7 (s, 2H), 6.6 (m, 3H),7.0 (m, 2H), 7.2 (m, 5H), 7.3 (m, 3H). MS: m/z 498.

The identity of Compound AO was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.5-1.7 (m, 6H), 2.3 (m, 4H), 2.4-3.1 (m, 8H), 3.4 (m,2H), 3.9 (t, 1H), 4.6 (s, 2H), 6.6-6.8 (m, 2H), 7.1-7.3 (m, 13H). MS:m/z 526.

5.15 Example 15 Synthesis of Compound AP

Compound AP was synthesized according to the method of Example 6 exceptthat ethyl 5-bromovalerate (commercially available from Sigma-Aldrich,St. Louis, Mo., www.sigma-aldrich.com) was used in place of ethyl3-bromopropionate. The purity of Compound AP was determined to begreater than 97% by HPLC analysis.

The identity of Compound AP was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): δ 1.7 (m, 7H), 2.3 (s, 3H), 2.4 (m, 2H), 2.7-2.9 (m,4H), 3.0 (s, 3H), 3.4-3.5 (m, 5H), 3.8 (m, 2H), 4.7 (s, 3H), 6.9 (t,1H), 7.15 (m, 2H), 7.3-7.4 (m, 12H). MS: m/z 597.

5.16 Example 16 Synthesis of Compound AQ

Compound AQ was synthesized according to the method of Example 6 exceptthat methyl bromoacetate (commercially available from Sigma-Aldrich, St.Louis, Mo., www.sigma-aldrich.com) was used in place of ethyl3-bromopropionate. The purity of Compound AQ was determined to begreater than 97% by HPLC analysis.

The identity of Compound AQ was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.9 (bs, 2H), 2.2 (m, 2H), 2.3 (bs, 3H), 2.4-2.6 (m,4H), 2.7 (m, 4H), 3.0 (bs, 3H), 3.7 (s, 3H), 4.1 (s, 2H), 4.7 (s, 2H),6.9 (m, 3H), 7.3 (m, 4H), 7.3-7.5 (m, 8H). MS: m/z 569.

5.17 Example 17 Synthesis of Compound AS

Compound AS was synthesized according to the method of Example 8 exceptthat ethyl 5-bromovalerate (commercially available from Sigma-Aldrich,St. Louis, Mo., www.sigma-aldrich.com) was used in place of ethyliodoacetate. The purity of Compound AS was determined to be greater than97% by HPLC analysis.

The identity of Compound AS was confirmed using ¹H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.3 (t, 3H), 1.7 (m, 8H), 2.3 (m, 2H), 2.5-2.9 (m,7H), 3.4 (m, 3H), 4.2 (q, 2H), 4.7 (s, 2H), 6.9 (m, 3H), 7.2-7.5 (m,12H). MS: m/z 579.

5.18 Example 18 Synthesis of Compound AU

Compound AU was synthesized according to the method of Example 9 exceptthat Compound AS (prepared according to the method of Example 17) wasused in place of Compound AH. The purity of Compound AU was determinedto be greater than 97% by HPLC analysis.

The identity of Compound AU was confirmed using 1H NMR and MS analysis.

¹H NMR (CDCl₃): δ 1.6-1.8 (m, 6H), 2.4 (m, 2H), 3.1-3.3 (m, 4H), 3.5 (m,6H), 3.7 (2H), 4.7 (s, 2H), 6.9 (m, 1H), 7.1 (m, 2H), 7.2-7.6 (m, 12H).MS: m/z 551.

5.19 Example 19 Synthesis of Compound AV

Compound AV was synthesized according to the method of Example 9 exceptthat Compound G, shown below:

was used in place of Compound AH.

Compound G was prepared according to the method of Example 8 except that3-iodopropionate (commercially available from Sigma-Aldrich, St. Louis,Mo., www.sigma-aldrich.com) was used in place of ethyl iodoacetate. Thepurity of Compound AV was determined to be greater than 97% by HPLCanalysis.

The identity of Compound AV was confirmed using ¹H NMR and MS analysis.

¹H NMR (CD₃OD): δ 2.0 (d, 2H), 2.7 (m, 4H), 3.0 (m, 2H), 3.2 (m, 2H),3.6 (m, 2H), 3.7 (m, 2H), 3.8 (m, 2H), 4.8 (s, 2H), 6.9 (m, 1H), 7.0 (m,2H), 7.3-7.5 (m, 12H). MS: m/z 523.

5.20 Example 20 Synthesis of Compound AI

A solution of Compound F (0.9 mmol) in 10 mL of DMF was added to a flaskcontaining 1.5 eq. of NaH (1.4 mmol) that had been flushed with argonand the resulting mixture was allowed to stir at room temperature forabout 15 min. 3-Bromopropionitrile (1.2 eq.) (commercially availablefrom Sigma-Aldrich, St. Louis, Mo., www.sigma-aldrich.com) was added tothe flask and the resulting mixture allowed to stirr at room temperaturefor 30 min. The mixture was then heated to 80° C. and allowed to stirfor about 12 h. The reaction mixture was then cooled to roomtemperature, diluted with 10 mL of ethylacetate, and washed with water(10 mL, 2 times). The organic layer was separated, dried (MgSO₄), andthe solvent removed under reduced pressure to provide a residue that waspurified using a silica gel column eluted with 10% triethylamine, 40%ethyl acetate, 50% hexanes to provide Compound AI.

5.21 Example 21 Synthesis of Compound AJ

Compound AJ was synthesized according to the method of Example 20 exceptthat 4-bromobutyronitrile (commercially available from Sigma-Aldrich,St. Louis, Mo., www.sigma-aldrich.com) was used in place of3-bromopropionitrile.

5.22 Example 22 μ- and ORL-1-Receptor-Binding Affinity Assays

5.22.1 Materials and Methods

ORL-1 Receptor Membrane Preparation

All reagents were from obtained from Sigma (St. Loius, Mo.) unless notedotherwise. Membranes from recombinant HEK-293 cells expressing the humanopioid receptor-like (ORL-1) receptor (Perkin Elmer, Boston, Mass.) wereprepared by lysing cells in ice-cold hypotonic buffer (2.5 mM MgCl₂, 50mM HEPES, pH 7.4) (10 mL/10 cm dish), followed by homogenization with atissue grinder/teflon pestle. Membranes were collected by centrifugationat 30,000×g for 15 min at 4° C., and pellets were resuspended inhypotonic buffer to a final concentration of 1-3 mg/mL. Proteinconcentrations were determined using the BioRad (Hercules, Calif.)protein assay reagent with bovine serum albumen as standard. Aliquots ofthe ORL-1 receptor membranes were stored at −80° C.

μ- and ORL-1-Receptor-Binding-Assay Procedures

Radioligand dose-displacement binding assays for ORL-1 and A receptorsused 0.1 nM [³H]-nociceptin or 0.2 n-M [³H]-diprenorphine (NEN, Boston,Mass.), respectively, with 5-20 mg membrane protein/well in a finalvolume of 500 ml binding buffer (10 mM MgCl₂, 1 mM EDTA, 5% DMSO, 50 mMHEPES, pH 7.4). Reactions were carried out in the absence or presence ofincreasing concentrations of unlabled nociceptin (American PeptideCompany, Sunnyvale, Calif.) or naloxone, for ORL-1 and μ, respectively.All reactions were conducted in 96-deep well polypropylene plates for1-2 h at room temperature. Binding reactions were terminated by rapidfiltration onto 96-well Unifilter GF/C filter plates (Packard, Meriden,Conn.) presoaked in 0.5% polyethylenimine using a 96-well tissueharvester (Brandel, Gaithersburg, Md.) followed performing by threefiltration washes with 500 mL of ice-cold binding buffer. Filter plateswere subsequently dried at 50° C. for 2-3 h. BetaScint scintillationcocktail (Wallac, Turku, Finland) was added (50 ml/well), and plateswere counted using a Packard Top-Count for 1 min/well. The data wereanalyzed using the one-site competition curve fitting functions inGraphPad PRISM v. 3.0 (San Diego, Calif.).

5.22.2 Results: μ-Receptor-Binding Assay

Generally, the lower the Ki value, the more effective the TriazospiroCompounds are at treating or preventing pain. Typically, the TriazaspiroCompounds have a Ki (nM) of about 300 or less for binding to μ-opioidreceptors. In one embodiment, the Triazaspiro Compounds have a Ki (nM)of about 100 or less. In another embodiment, the Triazaspiro Compoundshave a Ki (nM) of about 10 or less. In still another embodiment, theTriazaspiro Compounds have a Ki (nM) of about 1 or less. In stillanother embodiment, the Triazaspiro Compounds have a Ki (nM) of about0.1 or less.

For example, Compound AG, an illustrative Triazaspiro Compound, binds toμ-opioid receptors with a binding constant Ki of 2.9 nM. Accordingly,the above-disclosed assay indicates that the Triazaspiro Compounds wouldbe useful for treating or preventing pain in an animal.

5.22.3 Results: ORL-1-Receptor-Binding Assay

Generally, the lower the Ki value, the more effective the TriazospiroCompounds are at treating or preventing pain. Typically, the TriazaspiroCompounds have a Ki (nM) of about 10,000 or less for ORL-1 receptors. Inone embodiment, the Triazaspiro Compounds have a Ki (nM) of about 2000or less. In another embodiment, the Triazaspiro Compounds have a Ki (nM)of about 1000 or less. In still another embodiment, the TriazaspiroCompounds have a Ki (nM) of about 100 or less. In still anotherembodiment, the Triazaspiro Compounds have a Ki (nM) of about 10 orless.

For example, Compound AG, an illustrative Triazaspiro Compound, binds toORL-1 receptors with a binding constant Ki of 18 nM. Accordingly, theabove-disclosed assay indicates that the Triazaspiro Compounds would beuseful for treating or preventing pain in an animal.

5.23 Example 23 μ- and ORL-1-Opioid Receptor γS Functional Activity

5.23.1 Materials and Methods

[³⁵S]GTPgS functional assays were conducted using freshly thawed ORL-1or μ-receptor membranes, as appropriate. Assay reactions were preparedby sequentially adding the following reagents to binding buffer (100 mMNaCl, 10 mM MgCl₂, 20 mM HEPES, pH 7.4) on ice (final concentrationsindicated): membrane protein (0.066 mg/mL for ORL-1 receptor and 0.026mg/mL for it-receptor), saponin (10 mg/ml), GDP (3 mM) and [³⁵S]GTPgS(0.20 nM; NEN). The prepared membrane solution (190 mL/well) wastransferred to 96-shallow well polypropylene plates containing 10 mL of20× concentrated stock solutions of the agonist nociceptin prepared indimethyl sulfoxide (“DMSO”). Plates were incubated for 30 min at roomtemperature with shaking. Reactions were terminated by rapid filtrationonto 96-well Unifilter GF/B filter plates (Packard, Meriden, Conn.)using a 96-well tissue harvester (Brandel, Gaithersburg, Md.) followedby three filtration washes with 200 mL of ice-cold binding buffer (10 mMNaH₂PO₄, 10 mM Na₂HPO₄, pH 7.4). Filter plates were subsequently driedat 50° C. for 2-3 h. BetaScint scintillation cocktail (Wallac, Turku,Finland) was added (50 mL/well) and plates were counted using a PackardTop-Count for 1 min/well. Data were analyzed using the sigmoidaldose-response curve fitting functions in GraphPad PRISM, v. 3.0.

5.23.2 Results: μ-Receptor Function

μ GTP EC₅₀ is the concentration of a compound providing 50% of themaximal response for the compound at μ receptor. Triazaspiro Compoundstypically having a μ GTP EC₅₀ (nM) of about 5000 or less stimulate itopioid receptor function. In one embodiment, the Triazaspiro Compoundshave a μ GTP EC₅₀ (nM) of about 1000 or less. In still anotherembodiment, the Triazaspiro Compounds have a μ GTP EC₅₀ (nM) of about100 or less. In still another embodiment, the Triazaspiro Compounds havea μ GTP EC₅₀ (nM) of about 10 or less. In still another embodiment, theTriazaspiro Compounds have a μ GTP EC₅₀ (nM) of about 1 or less. Instill another embodiment, the Triazaspiro Compounds have a μ GTP EC₅₀(rAM) of about 0.1 or less.

μ GTP Emax % is the maximal effect elicited by a compound relative tothe effect elicited by [D-Ala2, N-methyl-Phe4, Gly-ol5]-enkephalin(“DAMGO”), a standard μ agonist. Generally, the μ GTP Emax (%) valuemeasures the efficacy of a compound to treat or prevent pain. Typicallythe Triazaspiro Compounds have a μ GTP Emax (%) of greater than 50%. Inone embodiment the Triazaspiro Compounds have a μ GTP Emax (%) ofgreater than 75%. In still another embodiment the Triazaspiro Compoundshave a μ GTP Emax (%) of greater than 88%. In still another embodimentthe Triazaspiro Compounds have a μ GTP Emax (%) of greater than 100%.

For example, Compound AG, an illustrative Triazaspiro Compound,stimulates μ-opioid-receptor function and exhibits a μ GTP EC₅₀ of 44 nMand a μ GTP Emax of 88%. Accordingly, this assay indicates thatTriazaspiro Compounds would be useful useful for treating or preventingpain in an animal.

5.23.3 Results: ORL-1-Receptor Function

ORL-1 GTP EC₅₀ is the concentration of a compound providing 50% of themaximal response for the compound at an ORL-1 receptor. TriazaspiroCompounds having a ORL-1 GTP EC₅₀ (nM) of about 10,000 or less stimulateORL-1 opioid-receptor function. In one embodiment, the TriazaspiroCompounds have a ORL-1 GTP EC₅₀ (nM) of about 1000 or less. In stillanother embodiment, the Triazaspiro Compounds have a ORL-1 GTP EC₅₀ (nM)of about 100 or less. In still another embodiment, the TriazaspiroCompounds have a ORL-1 GTP EC₅₀ (nM) of about 50 or less. In stillanother embodiment, the Triazaspiro Compounds have a ORL-1 GTP EC₅₀ (nM)of about 10 or less.

ORL-1 GTP Emax % is the maximal effect elicited by a compound relativeto the effect elicited by nociceptin, a standard ORL-1 agonist.Generally, the ORL-1 GTP Emax (%) value measures the efficacy of acompound to treat or prevent pain. Typically the Triazaspiro Compoundshave a ORL-1 GTP Emax (%) of greater than 50%. In one embodiment theTriazaspiro Compounds have a ORL-1 GTP Emax (%) of greater than 75%. Instill another embodiment the Triazaspiro Compounds have a ORL-1 GTP Emax(%) of greater than 88%. In still another embodiment the TriazaspiroCompounds have a ORL-1 GTP Emax (%) of greater than 100%.

For example, Compound AG, an illustrative Triazaspiro Compound,stimulates ORL-1 opioid-receptor function and exhibits a ORL-1 GTP EC₅₀of 71 nM and a ORL-1 GTP Emax of 95%. Accordingly, these assays indicatethat Triazaspiro Compounds would be useful for treating or preventingpain in an animal.

The present invention is not to be limited in scope by the specificembodiments disclosed in the examples which are intended asillustrations of a few aspects of the invention and any embodiments thatare functionally equivalent are within the scope of this invention.Indeed, various modifications of the invention in addition to thoseshown and described herein will become apparent to those skilled in theart and are intended to fall within the scope of the appended claims.

A number of references have been cited, the entire disclosures of whichare incorporated herein by reference.

1. A compound of formula (Ia):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is hydrogen,—COOH, —COOR³, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), —C(O)N(C₁-C₄ alkyl)(C₁-C₄alkyl), —CN, —N(H)S(O)₂(C₁-C₄ alkyl), or

R² is —COOH, —COOR³ or

R³ is —(C₁-C₆ alkyl), benzyl, phenyl, or —(C₃-C₆ cycloalkyl); n is 0when R¹ is hydrogen, and n is an integer ranging from 1 to 4 when R¹ isother than hydrogen; and m is an integer ranging from 0 to
 4. 2. Thecompound or pharmaceutically acceptable salt of the compound of claim 1,wherein R¹ is hydrogen and n is
 0. 3. The compound or pharmaceuticallyacceptable salt of the compound of claim 1, wherein R¹ is —COOH or—COOR³.
 4. The compound or pharmaceutically acceptable salt of thecompound of claim 1, wherein R¹ is —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl) or—C(O)N(C₁-C₄ alkyl)(C₁-C₄ alkyl).
 5. The compound or pharmaceuticallyacceptable salt of the compound of claim 1, wherein R¹ is —CN.
 6. Thecompound or pharmaceutically acceptable salt of the compound of claim 1,wherein R¹ is —N(H)S(O)₂(C₁-C₄ alkyl).
 7. The compound orpharmaceutically acceptable salt of the compound of claim 1, wherein R¹is


8. The compound or pharmaceutically acceptable salt of the compound ofclaim 1, wherein R² is —COOH or —COOR³.
 9. The compound orpharmaceutically acceptable salt of the compound of claim 1, wherein R²is


10. The compound or pharmaceutically acceptable salt of the compound ofclaim 1, where m is 0 or
 1. 11. The compound or pharmaceuticallyacceptable salt of the compound of claim 1, whererin m is
 0. 12. Thecompound of claim 1 having the structure:

or a pharmaceutically acceptable salt thereof.
 13. A compositioncomprising an effective amount of a compound or a pharmaceuticallyacceptable salt of the compound of claim 1 and a pharmaceuticallyacceptable carrier or excipient.
 14. The composition of claim 13,further comprising an opioid analgesic.
 15. The composition of claim 13,further comprising a non-opioid analgesic.
 16. The composition of claim13, further comprising an anti-emetic agent.
 17. A compound of formula(Ib):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is —COOH,—COOR³, —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), —C(O)N(C₁-C₄ alkyl)(C₁-C₄alkyl), —CN, —N(H)S(O)₂(C₁-C₄ alkyl), or

R² is —COOH,—COOR³, —C(O)N(CH₃)₂, or

R³ is —(C₁-C₆ alkyl), benzyl, phenyl, or —(C₃-C₆ cycloalkyl); n is aninteger ranging from 1 to 4; and m is an integer ranging from 0 to 4.18. The compound or a pharmaceutically acceptable salt of the compoundof claim 17, wherein R¹ is —COOH or —COOR³.
 19. The compound or apharmaceutically acceptable salt of the compound of claim 17, wherein R¹is —C(O)NH₂, —C(O)NH(C₁-C₄ alkyl) or —C(O)N(C₁-C₄ alkyl)(C₁-C₄ alkyl).20. The compound or a pharmaceutically acceptable salt of the compoundof claim 17, wherein R¹ is —CN.
 21. The compound or a pharmaceuticallyacceptable salt of the compound of claim 17, wherein R¹ is—N(H)S(O)₂(C₁-C₄ alkyl).
 22. The compound or a pharmaceuticallyacceptable salt of the compound of claim 17, wherein R¹ is


23. The compound or a pharmaceutically acceptable salt of the compoundof claim 17, wherein R² is —COOH or —COOR³.
 24. The compound orpharmaceutically acceptable salt of the compound of claim 17, whererin mis 0 or
 1. 25. The compound or pharmaceutcially acceptable salt of thecompound of claim 17, wherein m is
 0. 26. The compound or apharmaceutically acceptable salt of the compound of claim 17, wherein R²is —C(O)N(CH₃)₂.
 27. The compound or a pharmaceutically acceptable saltof the compound of claim 17, wherein R² is


28. The compound of claim 17 having the structure:

or a pharmaceutically acceptable salt thereof.
 29. A compositioncomprising an effective amount of a compound or a pharmaceuticallyacceptable salt of the compound of claim 17 and a pharmaceuticallyacceptable carrier or excipient.
 30. The composition of claim 29,further comprising an opioid analgesic.
 31. The composition of claim 29,further comprising a non-opioid analgesic.
 32. The composition of claim29, further comprising an anti-emetic agent.
 33. A compound of formula(Ic):

or a pharmaceutically acceptable salt thereof, wherein: R¹ is —COOH,—C(O)NH₂, —C(O)NH(C₁-C₄ alkyl), —C(O)N(C₁-C₄ alkyl)(C₁-C₄ alkyl),—N(H)S(O)₂(C₁-C₄ alkyl), or

n is an integer ranging from 1 to
 4. 34. The compound of claim 33,whererin n is
 2. 35. The compound of claim 33, wherein n is
 1. 36. Thecompound or a pharmaceutically acceptable salt of the compound of claim33, wherein R¹ is —COOH.
 37. The compound or a pharmaceuticallyacceptable salt of the compound of claim 33, wherein R¹ is —C(O)NH₂,—C(O)NH(C₁-C₄ alkyl) or —C(O)N(C₁-C₄ alkyl)(C₁-C₄ alkyl).
 38. Thecompound or a pharmaceutically acceptable salt of the compound of claim33, wherein R¹ is —N(H)S(O)₂(C₁-C₄ alkyl).
 39. The compound or apharmaceutically acceptable salt of the compound of claim 33, wherein R¹is


40. The compound of claim 33 selected from the group consisting of:

or a pharmaceutically acceptable salt thereof.
 41. A compositioncomprising an effective amount of a compound or a pharmaceuticallyacceptable salt of the compound of claim 33 and a pharmaceuticallyacceptable carrier or excipient.
 42. The composition of claim 41,further comprising an opioid analgesic.
 43. The composition of claim 41,further comprising a non-opioid analgesic.
 44. The composition of claim41, further comprising an anti-emetic agent.